Chapel and Haeney's Essentials of

Clinical Immunology (Jan 12,

2023)_(1119542383)_(Wiley-Blackwell)
7th Edition Spickett
Visit to download the full and correct content document:
https://ebookmass.com/product/chapel-and-haeneys-essentials-of-clinical-immunolog
y-jan-12-2023_1119542383_wiley-blackwell-7th-edition-spickett/
More products digital (pdf, epub, mobi) instant
download maybe you interests ...

Essentials of Biology 2023 7th Edition Sylvia S. Mader

https://ebookmass.com/product/essentials-of-biology-2023-7th-
edition-sylvia-s-mader/

Chronic Total Occlusions-A Guide to Recanalization, 3e

(Nov 29, 2023)_(1119517273)_(Wiley-Blackwell) Ron
Waksman

https://ebookmass.com/product/chronic-total-occlusions-a-guide-
to-recanalization-3e-nov-29-2023_1119517273_wiley-blackwell-ron-
waksman/

Clinical Immunology Nima Rezaei

https://ebookmass.com/product/clinical-immunology-nima-rezaei/

Contemporary Clinical Immunology and Serology (Pearson

Clinical Laboratory Science)

https://ebookmass.com/product/contemporary-clinical-immunology-
and-serology-pearson-clinical-laboratory-science/
Saunders 2022-2023 Clinical Judgment and Test-Taking
Strategies 7th Edition Linda Anne Silvestri

https://ebookmass.com/product/saunders-2022-2023-clinical-
judgment-and-test-taking-strategies-7th-edition-linda-anne-
silvestri/

Clinical Immunology: Principles and Practice 6th

Edition Robert R. Rich

https://ebookmass.com/product/clinical-immunology-principles-and-
practice-6th-edition-robert-r-rich/

Cochrane Handbook for Systematic Reviews of Diagnostic

Test Accuracy (Wiley Cochrane Series) (Jul 12,
2023)_(1119756162)_(Wiley-Blackwell) 1st Edition
Jonathan J. Deeks
https://ebookmass.com/product/cochrane-handbook-for-systematic-
reviews-of-diagnostic-test-accuracy-wiley-cochrane-series-
jul-12-2023_1119756162_wiley-blackwell-1st-edition-jonathan-j-
deeks/

Essentials of economics 7th Edition Garratt

https://ebookmass.com/product/essentials-of-economics-7th-
edition-garratt/

Essentials of Nursing Informatics, 7th Edition Virginia

Saba

https://ebookmass.com/product/essentials-of-nursing-
informatics-7th-edition-virginia-saba/
 CHAPELAND HAENEY ’ S ESSENTIALS OF
CLINICAL
IMMUNOLOGY
SIRAJ A. MISBAH I GAVIN P. SPICKETT I VIRGIL A.S. H . DALM
SEVENTH EDITION

ISTUDY
 Chapel and
Haeney’s
Essentials of
Clinical
Immunology
 Chapel and
Haeney’s
Essentials of
Clinical
Immunology
Seventh Edition
Siraj A. Misbah
MSc, FRCP, FRCPath
Consultant Clinical Immunologist, Honorary Senior Clinical Lecturer in Immunology
Oxford University Hospitals NHS Foundation Trust and University of Oxford
Oxford, UK

Gavin P. Spickett
MA, LLM, DPhil, FRCPath, FRCP, FRCP(E)
Retired Consultant Clinical Immunologist, Royal Victoria Infirmary
Newcastle upon Tyne, UK

Virgil A.S.H. Dalm

MD, PhD
Department of Internal Medicine, Division of Allergy & Clinical Immunology
Department of Immunology, Erasmus University Medical Center Rotterdam
Rotterdam, The Netherlands
This seventh edition first published 2022
© 2022 John Wiley & Sons Ltd
Edition History
Wiley-­Blackwell (6e, 2014)
All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted,
in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as
permitted by law. Advice on how to obtain permission to reuse material from this title is available at
http://www.wiley.com/go/permissions.
The right of Siraj A. Misbah, Gavin P. Spickett, and Virgil A.S.H. Dalm to be identified as the authors of
this work has been asserted in accordance with law.
Registered Offices
John Wiley & Sons, Inc., 111 River Street, Hoboken, NJ 07030, USA
John Wiley & Sons Ltd, The Atrium, Southern Gate, Chichester, West Sussex, PO19 8SQ, UK
Editorial Office
9600 Garsington Road, Oxford, OX4 2DQ, UK
For details of our global editorial offices, customer services, and more information about Wiley products
visit us at www.wiley.com.
Wiley also publishes its books in a variety of electronic formats and by print-­on-­demand. Some content
that appears in standard print versions of this book may not be available in other formats.
Limit of Liability/Disclaimer of Warranty
The contents of this work are intended to further general scientific research, understanding, and discussion
only and are not intended and should not be relied upon as recommending or promoting scientific method,
diagnosis, or treatment by physicians for any particular patient. In view of ongoing research, equipment
modifications, changes in governmental regulations, and the constant flow of information relating to the use
of medicines, equipment, and devices, the reader is urged to review and evaluate the information provided
in the package insert or instructions for each medicine, equipment, or device for, among other things,
any changes in the instructions or indication of usage and for added warnings and precautions. While the
publisher and authors have used their best efforts in preparing this work, they make no representations
or warranties with respect to the accuracy or completeness of the contents of this work and specifically
disclaim all warranties, including without limitation any implied warranties of merchantability or fitness for
a particular purpose. No warranty may be created or extended by sales representatives, written sales materials
or promotional statements for this work. The fact that an organization, website, or product is referred to
in this work as a citation and/or potential source of further information does not mean that the publisher
and authors endorse the information or services the organization, website, or product may provide or
recommendations it may make. This work is sold with the understanding that the publisher is not engaged
in rendering professional services. The advice and strategies contained herein may not be suitable for your
situation. You should consult with a specialist where appropriate. Further, readers should be aware that
websites listed in this work may have changed or disappeared between when this work was written and when
it is read. Neither the publisher nor authors shall be liable for any loss of profit or any other commercial
damages, including but not limited to special, incidental, consequential, or other damages.
Library of Congress Cataloging-­in-­Publication Data
Names: Misbah, Siraj A., author. | Spickett, Gavin, author. | Dalm, Virgil
A. S. H., author.
Title: Chapel and Haeney’s essentials of clinical immunology / Siraj
Misbah, Gavin Spickett, Virgil A.S.H. Dalm.
Other titles: Essentials of clinical immunology
Description: Seventh edition. | Hoboken, NJ : Wiley-Blackwell 2022. |
Preceded by Essentials of clinical immunology / Helen Chapel, Mansel
Haeney, Siraj Misbah, Neil Snowden. Sixth edition. 2014. | Includes
bibliographical references and index.
Identifiers: LCCN 2021061616 (print) | LCCN 2021061617 (ebook) | ISBN
9781119542384 (paperback) | ISBN 9781119542407 (adobe pdf ) | ISBN
9781119542421 (epub)
Subjects: MESH: Immune System Diseases | Immune System
Phenomena–physiology
Classification: LCC RC582 (print) | LCC RC582 (ebook) | NLM QW 740 | DDC
616.07/9–dc23/eng/20220110
LC record available at https://lccn.loc.gov/2021061616
LC ebook record available at https://lccn.loc.gov/2021061617
Cover Design: Wiley
Cover Image: © KATERYNA KON/GETTY IMAGES
Set in 10/12pt AGaramondPro by Straive, Pondicherry, India
Contents
Preface to the Seventh Edition vi
Preface to the First Edition vii
Key to Illustrations viii
About the Companion Website ix

1 Basic Components: Structure and Function 1

2 Infection 35
3 Immunodeficiency 55
4 Anaphylaxis and Allergy 87
5 Autoimmunity 106
6 Lymphoproliferative Disorders 122
7 Immune Manipulation 138
8 Transplantation 158
9 Kidney Diseases 172
10 Joints and Muscles 195
11 Skin Diseases 221
12 Eye Diseases 238
13 Chest Diseases 246
14 Gastrointestinal and Liver Diseases 263
15 Endocrinology and Diabetes 287
16 Non-malignant Haematological Diseases 298
17 Neuroimmunology 310
18 Immunological Diseases in Pregnancy 322
19 Techniques in Clinical Immunology 330

Appendix: Further Resources 349

Index 351
Preface to the Seventh Edition
The new authors have been honoured to take on the responsibility for this well-­respected textbook from
Professor Helen Chapel and Dr. Mansel Haeney, who originally created the book and have been heavily
involved in the previous six editions. In honour of their contributions both to the textbook and to develop-
ing and teaching clinical immunology, the book has now been renamed as Chapel and Haeney’s Essentials
of Clinical Immunology and we have tried not to deviate too far from their original aims.
The text has been updated as far as possible and all chapters have been revised. The original structure
and format have been retained. Many of the illustrations, even though quite old now, are still relevant to
illustrate key immunological principles. The Covid pandemic and the additional workload created, particu-
larly for Samras Johnson V, has meant that progress on the book has been slower than anticipated at the
initiation meeting pre-­pandemic. Covid has also led to significant advances in the clinical immunological
field and in particular it has kickstarted the clinical use of mRNA vaccines. Covid has produced its own
fascinating immunological problems and unexpected immunological responses to vaccines. Recognition of
‘long Covid’ has provided new impetus to investigate the causes of post-­infectious chronic fatigue. New
immunological therapies are being introduced at speed for all immunological diseases and gene manipula-
tion is going to have a huge influence on the management of genetic immunological diseases. Immunology
remains an exciting field that is involved in all areas of clinical medicine. We hope that we can convey this
excitement to the reader.
As always, the authors are grateful to their families and friends for their forebearance during the writing
process and also to many colleagues for direct and indirect help in updating the chapters. In particular, we
are most grateful to Dr Martin Barnardo, Consultant Clinical Scientist in Histocompatibility and
Immunogenetics for reviewing the chapter on Transplantation. The meticulous work of colleagues at LNS
Indexing is also gratefully acknowledged. We are grateful to James Watson, Anne Hunt and the very patient
team of editors at Wiley-­Blackwell. As most authors, we must apologise to them for being slow to respond
to their queries and to return proofs.

Siraj A. Misbah
Gavin P. Spickett
Virgil A.S.H. Dalm
Preface to the First Edition
Immunology is now a well-developed basic science and much is known of the normal physiology of the
immune system in both mice and men. The application of this knowledge to human pathology has lagged
behind research, and immunologists are often accused of practising a science which has little relevance to
clinical medicine. It is hoped that this book will point out to both medical students and practising clini-
cians that clinical immunology is a subject which is useful for the diagnosis and management of a great
number and variety of human disease.
We have written this book from a clinical point of view. Diseases are discussed by organ involvement,
and illustrative case histories are used to show the usefulness (or otherwise) of immunological investigations
in the management of these patients. While practising clinicians may find the case histories irksome, we
hope they will find the application of immunology illuminating and interesting. The student should gain
some perspective of clinical immunology from the case histories, which are selected for their relevance to
the topic we are discussing, as this is not a textbook of general medicine. We have pointed out those cases
in which the disease presented in an unusual way.
Those who have forgotten, or who need some revision of, basic immunological ideas will find them
condensed in Chapter 1. This chapter is not intended to supplant longer texts of basic immunology but
merely to provide a springboard for chapters which follow. Professor Andrew McMichael kindly contrib-
uted to this chapter and ensured that it was up-to-date. It is important that people who use and request
immunological tests should have some idea of their complexity, sensitivity, reliability and expense.
Students who are unfamiliar with immunological methods will find that Chapter 17 describes the tech-
niques involved.

Helen Chapel
Mansel Haeney
1984
Key to Illustrations
Throughout the illustrations standard forms have been used for commonly-occurring cells and pathways.
A key to these is given in the figure below.

USER GUIDE

Pre-B Pre-T B T Natural Plasma

lymphocyte lymphocyte lymphocyte lymphocyte killer cell cell

Macrophage Antigen-presenting cell Dendritic Mast cell Langerhans

(APC) cell cell

Basophil Eosinophil Neutrophil Monocyte Stem cell

About the Companion Website
The book is accompanied by a website:

www.wiley.com/go/misbah/immunology

The website contains:

• Interactive multiple-­choice questions
• Additional case studies and figures to test your knowledge
• Case studies from the book with additional figures for you to use for revision
• All figures from the book in PowerPoint format
• Useful website links

Scan this QR code to visit the companion website.

 1

CHAPTER 1

Basic Components:
Structure and
Function
Key topics
■■ 1.1 Introduction 2
■■ 1.2 Key molecules 2
■■ 1.2.1 Molecules recognized by immune systems 3
■■ 1.2.2 Recognition molecules 4
■■ 1.2.3 Accessory molecules 11
■■ 1.2.4 Effector molecules for immunity 11
■■ 1.2.5 Receptors for effector functions 13
■■ 1.2.6 Adhesion molecules 15
■■ 1.3 Functional basis of innate responses 15
■■ 1.3.1 Endothelial cells 17
■■ 1.3.2 Neutrophil polymorphonuclear leucocytes 17
■■ 1.3.3 Macrophages 18
■■ 1.3.4 Dendritic cells 18
■■ 1.3.5 Complement 20
■■ 1.3.6 Antibody-­dependent cell-­mediated cytotoxicity 23
■■ 1.3.7 Natural killer cells 23
■■ 1.3.8 Innate lymphoid cells 24
■■ 1.4 Functional basis of the adaptive immune responses 24
■■ 1.4.1 Antigen processing 24
■■ 1.4.2 T-­cell-­mediated responses 25
■■ 1.4.3 Antibody production 28
■■ 1.5 Physiological outcomes of immune responses 29
■■ 1.5.1 Killing of target cells (virally infected/tumour cells) 29
■■ 1.5.2 Direct functions of antibody 29
■■ 1.5.3 Indirect functions of antibody 29
■■ 1.5.4 Regulation 30
■■ 1.6 Tissue damage caused by the immune system 30
■■ 1.6.1 Inflammation: a brief overview 30
■■ 1.7 Organization of the immune system: an overview 33
■■ 1.8 Conclusions 34

Visit the companion website at www.wiley.com/go/misbah/immunology to

download cases on these topics.

Chapel and Haeney’s Essentials of Clinical Immunology, Seventh Edition. Edited by Siraj A. Misbah,
Gavin P. Spickett, and Virgil A.S.H. Dalm.
© 2022 John Wiley & Sons Ltd. Published 2022 by John Wiley & Sons Ltd.
2 / Chapter 1: Basic Components: Structure and Function

1.1 Introduction
The immune system evolved as a defence against infectious diseases. Individuals with markedly deficient immune
responses, if untreated, succumb to infections in early life. There is, therefore, a selective evolutionary pressure for a
really efficient immune system. Although innate systems are fast in response to pathogens, the evolution to adaptive
responses provided greater efficiency. However, a parallel evolution in pathogens means that all species, plants,
insects, fish, birds and mammals have continued to improve their defence mechanisms over millions of years, giving
rise to some redundancies as well as resulting in apparent complexity. The aim of this chapter is to provide an initial
description of the molecules involved, moving on to the role of each in the immune processes rather than the more
traditional sequence of anatomical structure, cellular composition and then molecular components. It is hoped that
this gives a sense of their relationship in terms of immediacy and dependency as well as the parallel evolution of the
two immune systems. An immune response consists of five parts:

• Recognition of material as foreign and dangerous.

• An early innate (non-­specific) response to this recognition.
• A slower specific response to a particular antigen, known as an adaptive response.
• Non-­specific augmentation of this response.
• A memory of specific immune responses, providing a quicker and larger response when that particular antigen is
encountered the second time.

Innate immunity, though phylogenetically older and important in terms of speed of response, is less efficient.
Humoral components (soluble molecules in the plasma) and cells in blood and tissues are involved. Such responses
are normally accompanied by inflammation and occur within a few hours of stimulation (Table 1.1).
Adaptive immune responses are also divided into humoral and cellular responses. Adaptive humoral responses
result in the generation of antibodies reactive with a particular antigen. Antibodies are proteins with similar structures,
known ­collectively as immunoglobulins (Ig). They can be transferred passively to another individual by injection of
serum. In contrast, only cells can transfer cellular immunity. Good examples of cellular immune responses are the
rejection of a graft by lymphoid cells as well as graft-­versus-­host disease, where viable transferred cells attack an
immunologically compromised recipient that is unable to fight back.
Antibody-­producing lymphocytes, which are dependent on the bone marrow, are known as B cells. In response to
antigen stimulation, B cells will mature to antibody-­secreting plasma cells. Cellular immune responses are dependent
on an intact thymus, so the lymphocytes responsible are known as thymus-­dependent (T) cells. The developmental
pathways of both cell types are fairly well established (Fig. 1.1).
The recognition phase is common to both adaptive and innate immunity. It involves professional cells, known
as classical dendritic cells (DCs), that recognize general pathogen features or specific antigenic molecules, process
the antigens and present antigen fragments to the other cells of the immune system, as well as initiating non-­specific
inflammation to the pathogen. In the effector phase, neutrophils and macrophages (innate immunity) and antibodies
and effector T lymphocytes (adaptive immunity) eliminate the antigen.
In terms of disease, like other organs, the immune system may fail (immunodeficiency), may become malignant (lymphoid
malignancies) or may produce aberrant responses (such as in autoimmunity or allergy). This chapter describes the normal
immune system in order to lay the basis for discussing these ways in which it can go wrong and so cause disease.

1.2 Key molecules
Many types of molecules play vital roles in both phases of
immune responses; some are shared by both the innate and the
adaptive systems. Antigens are substances that are recognized by
immune components. Detection molecules on innate cells rec-
ognize general patterns of ‘foreignness’ on non-­mammalian
cells, whereas those on adaptive cells are specific for a wide
range of very particular molecules or fragments of molecules.
Antibodies are not only the surface receptors of B cells (BCRs)
that recognize specific antigens, but, once the appropriate B
cells are activated and differentiate into plasma cells, antibodies
are also secreted into blood and body fluids in large quantities
to prevent that antigen from causing damage. T cells have
structurally similar receptors for recognizing antigens, known
as T-­cell receptors (TCRs). Major histocompatibility complex
(MHC) molecules provide a means of self-­recognition and also
play a fundamental role in T lymphocyte effector functions.
Effector mechanisms are often dependent on messages from
initiating or regulating cells; soluble mediators, which carry
messages between cells, are known as interleukins, cytokines
and chemokines.
 Chapter 1: Basic Components: Structure and Function / 3

Table 1.1 Components of innate and adaptive immunity

Features Innate Adaptive

Foreign molecules recognized Structures shared by microbes, recognized Wide range of very particular molecules or
as patterns (e.g. repeated glycoproteins), fragments of molecules on all types of extrinsic
PAMPs and modified self-­structures

Nature of recognition receptors Germline encoded – limited PRRs Somatic mutation results in a wide range of
specificities and affinities

Speed of response Immediate Time for cell movement and interaction between
cell types

Memory None Efficient

Humoral components Complement components Antibodies

Cellular components Dendritic cells, neutrophils, macrophages, Lymphocytes – T (Th1, Th2, Th17, Tregs), B
NK cells, NKT cells, B1 cells, epithelial
cells, mast cells

iNKT cells, γδ T cells

NK, natural killer; PAMPs, pathogen-­associated molecular patterns; pattern-­recognition receptors (PRRs); Tregs, regulatory T cells.

Lymphocyte development Peripheral effector cells

Myeloid
cell Th1

Th
Premyeloid Thymus T
cell Th2
Pre-T Thymocyte

Tc
Self reactive cells Natural
Pluripotential deleted T memory Killer cell
stem cell

Lymphocyte-
committed Secretory B
stem cell Bone
marrow

B
Pre-B Plasma cell

B memory

Pre-monocyte Monocyte Macrophage

Fig. 1.1 Development of different types of blood cells from a pluripotential stem cell in the bone marrow. The developmental pathway
for natural killer (NK) cells is shown separately because it is thought that NK cells may develop in both the thymus and the bone
marrow.

1.2.1 Molecules recognized by immune systems

signals’, due to pattern-­recognition receptors on DCs recogniz-
Foreign substances are recognized by both the innate and adap- ing conserved microbial structures directly, often repeated
tive systems, but in different ways, using different receptors polysaccharide molecules, known as pathogen-­associated
(see section 1.2.2). The innate system is activated by ‘danger molecular patterns. Toll-­like receptors (receptors that serve a
4 / Chapter 1: Basic Components: Structure and Function
similar function to toll receptors in drosophila) make up a large
family of non-­antigen-­specific receptors for a variety of indi-
vidual bacterial, viral and fungal components such as DNA,
lipoproteins and lipopolysaccharides. Activation of DCs by
binding to either of these detection receptors leads to inflam-
mation and subsequently activation of the adaptive system.
Phagocytic cells also recognize particular patterns associated
with potentially damaging materials, such as lipoproteins and
other charged molecules or peptides.
Traditionally, antigens have been defined as molecules that
interact with components of the adaptive system, i.e. T-­ and
B-­cell recognition receptors and antibody. An antigenic mole-
cule may have several antigenic determinants (epitopes); each
epitope can bind with an individual antibody, and a single
antigenic molecule can therefore provoke many antibody mol-
ecules with different binding sites. Some low-­molecular-­weight
molecules, called haptens, are unable to provoke an immune
response themselves, although they can react with existing anti-
bodies. Such substances need to be coupled to a carrier mole-
cule in order to have sufficient epitopes to be antigenic. For
some chemicals, such as drugs, the carrier may be a host (auto)
protein. The tertiary structure, as well as the amino acid
sequence, is important in determining antigenicity. Pure lipids
and nucleic acids are poor antigens, although they do activate
the innate system and can be inflammatory.
Antigens are conventionally divided into thymus-­dependent
and thymus-­independent antigens. Thymus-­dependent anti-
gens require T-­cell participation to provoke the production of
antibodies; most proteins are examples. Thymus-­independent
antigens require no T-­cell cooperation for antibody produc-
tion; they directly stimulate specific B lymphocytes by virtue of
their ability to cross-­link antigen receptors on the B-­cell sur-
face, produce predominantly IgM and IgG2 antibodies and
provoke poor immunological memory. Such antigens include
bacterial polysaccharides, found in bacterial cell walls.
Endotoxin, another thymus-­independent antigen, not only
causes specific B-­cell activation and antibody production, but
also acts as a stimulant for all B cells regardless of specificity.
Factors other than the intrinsic properties of the antigen
can also influence the quality of the immune response
(Table 1.2). Substances that improve an immune response to
a separate, often rather weak, antigen are known as adju-
vants. The use of adjuvants in humans, important in vac-
cines against infective agents and tumours, is discussed in
Chapter 7, section 7.3.2.
Superantigen is the name given to those foreign proteins
that are not specifically recognized by the adaptive system but
do activate large numbers of T cells regardless of specificity, via
direct action with an invariant part of the TCR (see Chapter 2,
section 2.4.2).
Self-­antigens are not recognized by DCs, so inflammation
and co-­stimulation of T cells (see section 1.4.1) is not induced.
There are mechanisms to control any aberrant adaptive
responses to self-­antigens, by prevention of the production of
specific receptors and regulation of the response if the immune
system is fooled into responding (see Chapter 5).
Table 1.2 Factors influencing the immune response
to an antigen, i.e. its immunogenicity
1 Nature of molecule:
Protein content
Size
Solubility
2 Dose:
Low doses provoke small amounts of antibody with
high affinity and restricted specificity
Moderate doses provoke large amounts of antibody but
mixed affinity and broad specificity
High doses provoke tolerance
3 Route of entry:
ID, IM, SC→regional lymph nodes
IV→spleen
Oral→Peyer’s patches
Inhalation→bronchial lymphoid tissue
4 Addition of substances with synergistic effects, e.g.
adjuvants
5 Genetic factors of recipient animal:
Species differences
Individual differences
ID, intradermal injection; IM, intramuscular injection; IV, intravenous
injection; SC, subcutaneous injection.
1.2.2 Recognition molecules
There are several sets of detection molecules on DCs
(Table 1.3): pattern-­recognition receptors (PRRs), such as Toll-­
like receptors, as well as chemotactic receptors and phagocytic
receptors. PRRs may be soluble or attached to cell membranes.
Mannan-­binding lectin is a protein that binds sugars on micro-
bial surfaces; if attached to a macrophage, it acts as a trigger for
phagocytosis and, if soluble, it activates the complement cas-
cade, resulting in opsonization. Others belonging to this family
are less well defined.
Toll-­like receptors (TLRs) are part of this family too and
are expressed either on the cell surface or intracellularly on
endosomal membranes (Table 1.4). These are evolutionarily
conserved proteins found on macrophages, DCs and neutro-
phils. At least ten different TLRs are found in humans, each
TLR recognizing a range of particular motifs on pathogens,
such as double-­stranded RNA of viruses (TLR3), lipopolysac-
charides of Gram-­negative bacterial cell walls (TLR4), flagellin
(TLR5) and bacterial DNA (TLR9), all highly conserved
motifs unique to microorganisms. Upon binding to their
ligands, TLRs induce signal transduction, via a complex cas-
cade of intracellular adaptor molecules and kinases, culminat-
ing in the induction of nuclear factor kappa B transcription
factor (NFκB)-­dependent gene expression and the induction
of proinflammatory cytokines (Fig. 1.2). The clinical conse-
quences of a defective TLR pathway are discussed in Chapter 3,
section 3.4.1 (see Box 1.1 in this chapter also).
Inflammasomes are a complex of intracellular proteins that
are assembled in response to sensing pathogen-­associated
 Chapter 1: Basic Components: Structure and Function / 5

Table 1.3 Markers on dendritic cells

Immature dendritic cells Mature myeloid dendritic cells

Function Antigen capture Antigen presentation to immature

T cells for specific differentiation
Co-­stimulatory molecule expression, Absent or low ++
e.g. CD80, CD86
Adhesion molecules, e.g. ICAM-­1 Absent or low ++
Cytokine receptors, e.g. IL-­12R Absent or low ++
Pattern-­recognition receptors, e.g. ++ –
mannose receptor
MHC class II:
Turnover Very rapid Persist >100 h
Density Reduced (approx. 1 × 106) Very high (approx. 7 × 106)

ICAM-­1, Intercellular adhesion molecule-­1; IL, interleukin; MHC, major histocompatibility complex.

Table 1.4 Location and ligands for toll-­like receptors (TLRs)

TLR Location Ligand

TLR1, TLR2 Cell surface Bacterial lipopeptides, peptidoglycan

TLR3 Endosomal membrane ds viral RNA
TLR4 Cell surface Lipopolysaccharide
TLR6 Cell surface Bacterial lipopeptides
TLR7 Endosomal membrane ssRNA
TLR8 Endosomal membrane ssRNA
TLR9 Endosome CpG DNA

Viruses Fig. 1.2 Sequential cellular events induced by

Ligands
engagement of Toll-­like receptors on dendritic cells,
Lipopolysaccharide Gram-negative
neutrophils and macrophages by microbial ligands.
(LPS) bacteria
or or IKB, inhibitor kappa B; IRAK, interleukin-­1 receptor-­
associated kinase; MAPK, mitogen-­activated protein
kinase; TRAF, TNF receptor-­associated factor.
Toll-like
receptors
(TLRs) Myd 88
(adaptor
protein)

Family of IRAK enzymes

Signalling
pathways
TRAF

Inactivation Induction of
of IKB MAPK kinases
Outcomes
Translocation of NFκB
Pro-inflammatory
cytokine secretion
Activation of Activation of genes
adaptive immunity in the nucleus
6 / Chapter 1: Basic Components: Structure and Function

Box 1.1 Clinical consequences of a or chain or chain

defective Toll-­like receptor pathway
CD3 Variable
In humans, deficiency of IRAK-­4 (interleukin-­1 region

receptor-­associated kinase) or MyDD88, key intracellular

­molecules responsible for TLR signal transduction Constant
region
(Fig. 1.2), is associated with recurrent pyogenic bacterial
infections accompanied by failure to mount an appropriate
acute-­phase response (see Chapter 3, Case 3.6). Plasma
Mice lacking TLR4 are exceptionally susceptible to membrane
ZAP70 p56lck p59fyn
infection with Gram-­negative bacteria.
Fig. 1.4 Diagram of the structure of the T-­cell receptor (TCR). The
variable regions of the alpha (ɑ) and beta (β) chains make up the
T idiotype, i.e. antigen/peptide-­binding region. The TCR is closely
LPS associated on the cell surface with the CD3 protein that is
essential for activation.
TLR-4

intestine and other microbial-­rich surfaces. CD1 present lipids

to the immune cells of the gut in particular, namely non-­
MHC-­restricted natural killer (NK) T cells and γδ T cells in
the epithelium.
NF-κB
activation
Antigenic epitopes, having been processed by DCs, are rec-
Recruitment
of Caspase
ognized by cells of the adaptive system by means of specific
proteaser receptors. Each T cell, like B cells, is pre-­committed to a given
IL-1B epitope. It recognizes this by one of two types of TCRs, depend-
TL-18
Assembly of
ing on the cell’s lineage and thus its effector function. T cells
NLRP3
inflammassre
have either αβTCR – a heterodimer of alpha (α) and beta β)
Actiotion of 3 enes
for NLR 3, IL-1
chains – or γδTCR – a heterodimer of gamma γ and delta (δ)
chains. αPTCR cells predominate in adults, although 10% of
T cells in epithelial structures are of the γδTCR type. In either
Nucleus case, TCRs are associated with several transmembrane proteins
that make up the cluster differentiation 3 (CD3) molecule
Fig. 1.3 Schematic representation of NLRP3 inflammasome
(Fig. 1.4), to make the CD3–TCR complex responsible for
activation. taking the antigen recognition signal inside the cell (signal
transduction). Signal transduction requires a group of intracel-
lular tyrosine kinases (designated p56 lck, p59 fyn, ZAP 70) to
join with the cytosolic tails of the CD3–TCR complex and
molecular patterns (PAMPs) and danger-­associated molecular become phosphorylated. Nearby accessory molecules, CD2,
patterns (DAMPs), derived from external pathogens or dam- LFA-­1, CD4 and CD8, are responsible for increased adhesion
aged host cells (Fig. 1.3). A typical inflammasome is built (see section 1.2.6), but are not actually involved in recognizing
around a scaffold containing NOD-­like receptors (NLRs). An presented antigenic fragments.
example of a clinically relevant, well-­characterized inflammas- The genes for TCR chains are on different chromosomes: β
ome is NLRP3, also known as cryopyrin. NLRP3 plays a key and γ on chromosome α7 and α and δ on chromosome 14.
role in innate immunity by orchestrating the release of proin- Each of the four chains is made up of a variable and a constant
flammatory cytokines, in particular interleukin (IL)-­1 and IL-­ domain. The variable regions are numerous (although less so
18. Mutations in the gene encoding cryopyrin are associated than immunoglobulin-­variable genes); they are joined by D
with a number of hereditary periodic fever syndromes (see and J region genes to the invariant (constant) gene by recombi-
Chapter 10), which are effectively treated by inhibitors of the nases, RAG1 and RAG2, the same enzymes used for making anti-
IL-­1 pathway (e.g. Anakinra). gen receptors on B cells and antibodies (section 1.4.1). The
CD1 molecules are invariant proteins (MHC-­like and diversity of T-­cell antige0.n receptors is achieved in a similar
associated with β2-­microglobulin – see later) that are present on way for immunoglobulin, although TCRs are less diverse since
DCs and epithelial cells. CD1 combine with lipids, which are somatic mutation is not involved; perhaps the risk of ‘self-­
poor antigens and not usually well presented to the adaptive recognition’ would be too great. The diversity of antigen bind-
immune system, and so act as recognition molecules for the ing is dependent on the large number of V genes and the way

in which these may be combined with different D and J genes
to provide different V domain genes. The similarities between
TCRs and BCRs led to the suggestion that the genes evolved
from the same parent gene and both are members of a ‘supergene’
family. Unlike immunoglobulin, TCRs are not secreted and are
not independent effector molecules.
A particular TCR complex recognizes a processed antigenic
peptide in the context of MHC class I or II antigens (sec-
tion 1.4.1), depending on the type of T cell; helper T cells
recognize class II with antigen, and this process is enhanced by
the surface accessory protein CD4 (see later) and intracellular
signals. Cytotoxic T cells (CTL/Tc) recognize antigens with
class I (see section 1.3.1) and use CD8 accessory molecules for
increased binding and signalling. Since the number of variable
genes available to TCRs is more limited, reactions with antigen
might not be sufficient if it were not for the increased binding
resulting from these accessory mechanisms. Recognition of
processed antigen alone is not enough to activate T cells.
Additional signals, through soluble cytokines (interleukins),
are needed; some of these are generated during ‘antigen pro-
cessing’ (see section 1.4.1).
MHC molecules were originally known as ‘histocompati-
bility antigens’ because of the vigorous reactions they provoked
during mismatched organ transplantation. However, these
molecules are known to play a fundamental role in immunity
by presenting antigenic peptides to T cells. Histocompatibility
antigens in humans (known as human leucocyte antigens,
HLAs) are synonymous with the MHC molecules. MHC mol-
ecules are cell-­surface glycoproteins of two basic types: class I
and class II (Fig. 1.5). They exhibit extensive genetic polymor-
phism with multiple alleles at each locus. As a result, genetic
variability between individuals is very great and most unrelated
individuals possess different MHC (HLA) molecules. This
means that it is very difficult to obtain perfect HLA matches
between unrelated persons for transplantation (see Chapter 8).
The extensive polymorphism in MHC molecules is best
explained by the need of the immune system to cope with an
Peptide binding groove
α1 α2 α1 β1
CHO CHO
CHO
CHO
s s s s
β 2m s s s s
α3 α2 β2
Plasma
membrane
Class I Class II
Fig. 1.5 Diagrammatic representation of major histocompatibility
complex (MHC) class I and class II antigens. CHO, carbohydrate
side chain; β2m, β2-­microglobulin.
Chapter 1: Basic Components: Structure and Function / 7
ever-­increasing range of pathogens adept at evading immune
responses (see Chapter 2).
The TCR of an individual T cell will only recognize antigen
as a complex of antigenic peptide and self-­MHC (Fig. 1.6).
This process of dual recognition of peptide and MHC mole-
cule is known as MHC restriction, since the MHC molecule
restricts the ability of the T cell to recognize antigen (Fig. 1.6).
The importance of MHC restriction in the immune response
was recognized by the award of the Nobel Prize in Medicine to
Peter Doherty and Rolf Zinkernagel, who found that virus-­
specific CTLs would only kill cells of the same particular allelic
form of MHC molecule.
MHC class I antigens are subdivided into three groups: A,
B and C. Each group is controlled by a different gene locus
within the MHC region on chromosome 6 (Fig. 1.7) in
humans (different in mice). The products of the genes at all
three loci are chemically similar. All MHC class I antigens (see
Fig. 1.5) are made up of an a heavy chain, controlled by a gene
in the relevant MHC locus, associated with a smaller chain
called β2-­microglobulin, controlled by a gene on chromosome
12. The differences between individual MHC class I antigens
are due to variations in the a chains; the β2-­microglobulin com-
ponent is constant. The detailed structure of class I antigens
was determined by X-­ray crystallography. This shows that
small antigenic peptides (approx. nine amino acids long) can
be tightly bound to a groove produced by the pairing of the
two extracellular domains (a1 and a2) of the a chain. The affin-
ity (tightness of binding) of individual peptide binding depends on
the nature and shape of the groove, and accounts for the MHC
restriction mentioned earlier.
MHC class II antigens have two heavy chains, α and β,
both coded for by genes in the MHC region of chromosome 6.
The detailed structure of MHC class II antigens was also
APC APC APC
MHC MHC MHC
type a type b type a
Ag Ag Ag
P P Q
TCR TCR TCR
T cell T cell T cell
RESPONDS NO RESPONSE NO RESPONSE
(i) (ii) (iii)
Fig. 1.6 Major histocompatibility complex (MHC) restriction of
antigen recognition by T cells. T cells specific for a particular
peptide and a particular MHC allele (i) will not respond if the
same peptide were to be presented by a different MHC molecule
as in (ii) or as in (iii) if the T cell were to encounter a different
peptide. APC, antigen-­presenting cell; TCR, T-­cell receptor.
8 / Chapter 1: Basic Components: Structure and Function

Presentation of Presentation of
Cell endogenous/viral antigens exogenous antigens
B C by MHC class I molecules by MHC class II molecules
membrane DR
DQ
DP A

Class III
Class II DQ Bf C4B Class I Vesicle
DP DR C2 C4A TNF B C A

Golgi Class II mRNA

endoplasmic
Centromere reticulum
Complex
with MHC I
Class I
mRNA
Endoplasmic Nucleus
Chromosome 6
reticulum Vesicle

Fig. 1.7 Major histocompatibility complex on chromosome 6; Viral antigen

class III antigens are complement components. TNF, tumour complexed
necrosis factor. with TAP

Viral
determined by X-­ray crystallography. It has a folded structure antigenic Viral
Endosome
peptide mRNA
similar to class I antigens, with the peptide-­binding groove
found between the α and β chains (see Fig. 1.5). Whereas most Viral DNA
Invariant
nucleated cells express class I molecules, expression of class II chain is cleaved on fusion
molecules is restricted to a few cell types: DCs, B lymphocytes, to enable class II molecules
activated T cells, macrophages, inflamed vascular endothelium Viral DNA to bind antigen in the groove

and some epithelial cells. However, other cells (e.g. thyroid,

Viral antigen/autoantigen Processed exogenous antigen
pancreas, gut epithelium) can be induced to express class II
molecules under the influence of interferon (IFN)-­γ released MHC class I molecule MHC class II molecule

during inflammation. In humans, there are three groups of
variable class II antigens: the loci are known as HLA-­DP
HLA-­DQ and HLA-­DR.
In practical terms, there are different mechanisms by which
antigens in different intracellular compartments can be cap-
tured and presented to CD4+ or CD8+ T cells (Fig. 1.8).
Endogenous antigens (including viral antigens that have
infected host cells) are processed by the endoplasmic reticulum
and presented by MHC class I-­bearing cells exclusively to
CD8+ T cells. Prior to presentation on the cell surface, endog-
enous antigens are broken down into short peptides, which are
then actively transported from the cytoplasm to endoplasmic
reticulum by proteins. These proteins act as a shuttle and are so
named ‘transporters associated with antigen processing’ (TAP-­1
and TAP-­2). TAP proteins (also coded in the MHC class II
region) deliver peptides to MHC class I molecules in the endo-
plasmic reticulum, from whence the complex of MHC and
peptide is delivered to the cell surface. Mutations that affect
function in either TAP gene prevent surface expression of
MHC class I molecules.
In contrast, exogenous antigens are processed by the lysoso-
mal route and presented by MHC class II antigens to CD4+ T
cells (Fig. 1.8). As with MHC class I molecules, newly synthe-
sized MHC class II molecules are held in the endoplasmic reticu-
lum until they are ready to be transported to the cell surface.
While in the endoplasmic reticulum, class II molecules are
prevented from binding to peptides in the lumen by a p ­ rotein
known as MHC class II-­associated invariant chain. The invariant
TAP (transporters associated Invariant chain protects
with antigen processing) antigen binding groove
Fig. 1.8 Different routes of antigen presentation, depending on
the nature of the antigen. MHC, major histocompatibility complex.
chain also directs delivery of class II molecules to the endosomal
compartment, where exogenous antigens are processed and made
available for binding to class II molecules.
The MHC class III region (see Fig. 1.7) contains genes
encoding proteins that are involved in the complement system
(see section 1.4.1): the early components C4 and C2 of the
classical pathway and factor B of the alternative pathway. Some
inflammatory proteins, e.g. tumour necrosis factor (TNF), are
also encoded in adjacent areas. Invariant MHC-­like proteins,
such as CD1 lipid-­recognition receptors (see earlier), are not
coded for on chromosome 6, despite being associated with
β2-­microglobulin.
In contrast to TCRs, the antigen BCRs are surface-­bound
immunoglobulin molecules that can be secreted as soluble
molecules. As with TCRs, they have predetermined specificity
for epitopes and are therefore extremely diverse. The immune
system has to be capable of recognizing all pathogens, past and
future. Such diversity is provided by the way in which all three
types of molecules, TCR, BCR and antibody, are produced.
The basic structure of the immunoglobulin molecule is
shown in Fig. 1.9. It has a four-­chain structure: two identical
 Chapter 1: Basic Components: Structure and Function / 9

VH
The amino (N) terminal domains of the heavy and light
N
terminal chains include the antigen-­binding site. The amino acid
sequences of these N-­terminal domains vary between different
Hinge
region CH1 antibody molecules and are known as variable (V) regions.
VL Most of these differences reside in three hypervariable areas of
CH3 CH2 the molecule, each only 6–10 amino acid residues long. In the
S folded molecule, these hypervariable regions in each heavy and
CL
S
light chain come together to form, with their counterparts on
C terminal
the other pair of heavy and light chains, the antigen-­binding
site (Fig. 1.9). The structure of this part of the antibody mole-
cule is unique to that molecule and is known as the idiotypic
Fc VL
determinant. In any individual, approximately 106–107 differ-
ent antibody molecules could be made up by 103 different
heavy chain variable regions associating with 103 different light
Fab
VH
chain variable regions, though there are even more epitopes
due to further variation during the later processing (see
section 1.4.1).
Fig. 1.9 Basic structure of an immunoglobulin molecule.
Domains are held in shape by disulfide bonds, though only one
The part of the immunoglobulin chain next to the V region
is shown. C1-­3, constant domain of a heavy chain; CL, constant in either heavy or light chains is the constant (C) region; this is
domain of a light chain; =S=, disulfide bond; VH, variable domain made up of one domain in a light chain (CL) and three or four
of a heavy chain; VL, variable domain of a light chain. in a heavy chain (CH) (Fig. 1.9). There are two alternative
types of CL chain, known as kappa (κ) and lambda (λ); an anti-
body molecule has either two κ or two λ light chains, never one
of each. Of all the antibodies in a human individual, roughly
IgM
60% contain κ and 40% contain λ light chains. There are no
J chain known differences in the functional properties between κ and
IgM IgM
λ light chains. In contrast, there are several possible different
types of CH domain, each with important functional differ-
ences (Table 1.5). The heavy chains determine the isotype of
the antibody and the ultimate physiological function of the
particular antibody molecule. Once the antigen-­binding site
IgM IgM has reacted with its antigen, the molecule undergoes a change
in the conformation of its heavy chains in order to take part in
Fig. 1.10 Schematic representation of immunoglobulin (Ig)M effector reactions, depending on the isotype of the molecule.
pentamer (MW 800 kDA). The processes by which the components of this supergene
family are produced are identical for TCR and BCR and are
known as recombination. Immunoglobulin production,
J chain
whether for BCR or antibody production, is the same initially.
IgA IgA As for the TCR, the genes for the different chains in a BCR are
carried on different chromosomes (Fig. 1.12). Like those cod-
ing for other macromolecules, the genes are broken up into
coding segments (exons) with intervening silent segments
(introns). The heavy chain gene set on chromosome 14 is made
Secretory piece
up of small groups of exons representing the constant regions
Fig. 1.11 Schematic representation of dimeric secretory immuno- of the heavy chains – e.g. mu (μ) chain – and a very large num-
globulin (Ig)A (MW 385 kDA). ber of V region genes, perhaps as many as 103. Between the V
and C genes are two small sets of exons, D and J (Fig. 1.12). In
a single B cell, one V region gene is selected, joined to one D
heavy (H) chains (mol. wt. 50 kDa) and two identical light (L) and J on the same chromosome; the VDJ product is then
chains (mol. wt. 25 kDa). Each chain is made up of domains of joined at the level of RNA processing to Cμ when the B cell is
about 110 amino acids held together in a loop by a disulphide making IgM. The cell can make IgG by omitting the Cμ and
bond between two cysteine residues in the chain. The domains joining VDJ to a Cγ. Thus, the cell can make IgM, IgD and
have the same basic structure and many areas of similarity in IgG/A/E in sequence, while still using the same variable region.
their amino acid sequences. The heavy chains determine the The same enzymes are used for the TCRs, and coded for by two
isotype of the immunoglobulin, resulting in pentameric IgM recombination-­activating genes controlling VDJ gene recombina-
(Fig. 1.10), dimeric IgA (Fig. 1.11) or monomeric IgG. tion: RAG1 and RAG2. Disruption of the RAG1 or RAG2
10 / Chapter 1: Basic Components: Structure and Function

Table 1.5 Immunoglobulin classes and their functions

Heavy Serum Complement Placental Reaction with

Isotype chain concentration* Main function fixation† passage Fc receptors‡
IgM μ 0.5–2.0 Neutralization and +++ – L
opsonization

IgG1 γ1 5.0–12.0 Opsonization +++ ++ M, N, P, L, E

IgG2 γ2 2.0–6.0 + ± P, L

IgG3 γ3 0.5–1.0 Opsonization +++ ++ M, N, P, L, E

IgG4 γ4 0.1–1.0 – + N, L, P

IgA1 α1 0.5–3.0 Neutralization at – – M, N

mucosal
surfaces

IgA2 α2 0.0–0.2 – – –

IgD δ Trace Lymphocyte – – –

membrane
receptor

IgE εΣ Trace Mast cell attachment – – B, E, L

* Normal adult range in g/L.
†
Classical pathway.
‡
Fc receptors on: basophils/mast cells, B; eosinophils, E; lymphocytes, L; macrophages, M; neutrophils, N; platelets, P.

function in infants who have mutations in these genes causes
profound immune deficiency, characterized by absent mature
B and T cells, as neither TCRs or BCRs can be produced. On
a different chromosome (either chromosome 22 for λ chains or
chromosome 2 for κ chains) in the same cell, a V gene is joined
to a J gene (there is no D on the light chain) and then the VJ
product is joined at the RNA level to the Cκ or Cλ (Fig. 1.12).
The wide diversity of antigen binding is dependent on the
large number of V genes and the way in which these may be
combined with different D and J genes to provide different rear-
ranged VDJ gene segments. Once V, D and J rearrangement has
taken place to produce a functional immunoglobulin molecule,
further V region variations are introduced only at a much later
stage, when antibodies rather than BCRs are produced by the
process of somatic mutation in germinal centres.
Natural killer cells also have recognition molecules.
These cells are important in killing virally infected cells and
tumour cells. They have to be able to recognize these targets
and distinguish them from normal cells. They recognize and
kill cells that have reduced or absent MHC class I, using two
kinds of receptors – inhibitory (KIR) and activating (KAR) –
to estimate the extent of MHC expression. They also have
one type of Fc IgG (Fcγ) receptor, that for low-­affinity binding
of IgG antibodies, and so NK cells are able to kill some cells
with large amounts of antibody on their surfaces. Further
subsets of NK-­like cells that contribute to innate immunity

Fig. 1.12 Immunoglobulin genes (see Chromosome ‘Silent’ area = Intron

text for explanation).
(V)n D J C Cδ Cγ3 Cγ1 Cα 1 Cγ2 Cγ4 Cε Cα 2
14 H

The product is VHC, i.e. an IgM heavy

VDJC chain with a particular variable region Final
product
(V)n J Cκ IgMκ
2 κ or
IgMλ
VJCκ
(V)n J Cλ
22 λ

VJCλ
 Chapter 1: Basic Components: Structure and Function / 11

include NKT cells and invariant NKT cells (section 1.3.6); (see section 1.4.3). Adhesion molecules, for binding leucocytes
these are thought to be particularly important in tumour (both lymphocytes and polymorphonuclear leucocytes) to
immunology (section 1.5.1). endothelial cells and tissue matrix cells, are considered in section
The major purpose of the complement pathways is to pro- 1.2.6. On B cells, such molecules include CD40 (ligand for
vide a means of removing or destroying antigens, regardless of CD40L, now named CD154; see Chapter 3, Case 3.2), B-­7-­1
whether or not these are coated with antibody. This requires and B7-­2 (ligands for CD28).
that complement components recognize damaging material such
as immune complexes (antigen combined with antibodies) or 1.2.4 Effector molecules for immunity
foreign antigens. The complement pathways are discussed in
more detail in section 1.3.5. There are humoral and cellular effector molecules in both
the innate and the adaptive immune systems (Table 1.6).
Several of the same mechanisms are used in both types of
1.2.3 Accessory molecules immune responses, especially in killing of target cells, sug-
The binding of a processed antigen–MHC class II complex on gesting that the evolution of immune responses has been
an antigen-­presenting cell (APC) to the corresponding TCR conservative in terms of genes, though with much redun-
provides an insufficient signal for T-­cell activation; the binding dancy to ensure the life-­preserving nature of the immune
of accessory molecules on the two cell surfaces provides systems in the face of rapid evolution of pathogenic microbes.
additional stimuli. Accessory molecules are lymphocyte surface
proteins, distinct from the antigen-­binding complexes, which Antibodies
are necessary for efficient binding, signalling and homing.
Accessory molecules are invariant, non-­polymorphic proteins. Each Antibodies are the best-­described effector mechanisms in
accessory molecule has a particular ligand – a corresponding adaptive immunity. They are the effector arm of B cells and
protein to which it binds. These ligands are present on all cells are secreted as soluble molecules by plasma cells in large
that require close adhesion for functioning; for example, there
are those on T cells for each of the many cell types that can Table 1.6 Effector molecules in immunity
activate or respond to T cells (APCs, endothelial cells, etc.);
Innate Adaptive
similar ligands are present on B cells for efficiency of T-­cell help
as well as stimulation by follicular DCs. Humoral Complement Specific antibodies for
components for opsonization and
There are several families of accessory molecules, but the most
opsonization or phagocytosis or lysis
important appear to be the immunoglobulin supergene fam- lysis with complement
ily of adhesion molecules, which derives its name from the
fact that its members contain a common immunoglobulin-­like Cellular Perforin in NK cells Perforin in cytolytic
structure. Members of the family strengthen the interaction creates pores in (CD8) T cells creates
target cell pores in specific target
between APCs and T cells (Fig. 1.13); those on T cells include
membranes cell membranes, allowing
CD4, CD8, CD28, CTLA-­4, CD45R, CD2 and lymphocyte entry of granzymes to
function antigen 1 (LFA-­1). For interaction with B cells, cause apoptosis
CD40 ligand and ICOS are important for class switching
NKT cells induce
apoptosis by perforin
Cell membrane Cell membrane production

TCR
Granzymes in NK
MHC class I or II cells induce
CD4 or CD8
apoptosis in target
(ICAM-1) CD54 (LFA-1) CD11a/CD18 cells
(LFA-3) CD58 CD2
Lysosomes in
(B7.1) CD80 CTLA-4 phagocytic
vacuoles result in
(B7.2) CD86 CD28
death of ingested
CD40 CD40L microbes

APC/virus infected T cell Preformed

target cell histamine and
related vasoactive
Fig. 1.13 Diagrammatic representation of adhesion molecules on substances as well
T cells and their ligands on antigen-­presenting cells (APCs)/ as leukotrienes in
virus-­infected target cells. MHC, major histocompatibility mast cells
complex; TCR, T-­cell receptor. NK, natural killer
Another random document with
no related content on Scribd:
throws light on a much contested subject; namely, the nature of the
so-called "intertentacular organ" (i, Fig. 234, p. 469), described so
long ago as 1837 by Farre,[573] but looked for in vain by the majority
of later observers.

The failure to find this organ, even in species which possess it, in
certain individuals, according to Farre's statements, is now
satisfactorily explained by M. Prouho, who shows that while it is
absent in a large number of polypides, it is normally present in those
individuals which possess an ovary, and in those only; and that its
primary function is that of an oviduct.

The intertentacular organ is an unpaired ciliated tube, which is

situated between the two tentacles which are nearest to the
ganglion. In the retracted condition of the polypide, it opens from the
body-cavity into the tentacle-sheath; and in the expanded condition,
directly to the exterior.

In the remarkable case of Alcyonidium duplex, each zooecium

normally possesses two sexual polypides. The first of these
produces a testis and then becomes a "brown body." The second is
meanwhile developed, and produces an ovary and an intertentacular
organ, a structure which was not present in the male polypide. The
eggs pass through the intertentacular organ into the tentacle-sheath,
and attach themselves to the diaphragm (d, Fig. 234), where they
remain during their development.

Although the intertentacular organ has been found by Prouho in

female polypides only, it would perhaps be going too far to assert
that it is confined to polypides of that sex. Hincks[574] has observed
the passage of spermatozoa in enormous numbers through the
organ, although it may be noted that there is no sufficient proof that
eggs were not present as well in these zooecia. It further appears
that in some cases waste matters may be removed from the body-
cavity through the same passage.
It may be presumed that the egg is normally fertilised by a
spermatozoon, although this is at present largely a matter of
inference. It is believed by Joliet[575] that fertilisation is reciprocal,
although Prouho has come to the opposite conclusion. Joliet has,
however, very justly pointed out that the enormous number of
spermatozoa developed by a single individual would be
disproportionately large, if their function were merely to fertilise the
ovum in the same zooecium. According to his view, the egg is
fertilised by a spermatozoon after it has passed into the tentacle-
sheath or ovicell, or some other place where it is in free
communication with the outside water.

Development and Affinities.—Few parts of the history of the

Polyzoa are more fascinating than that which deals with their
development; and it is probable that no other is capable of giving so
much insight into the affinities of the several groups to one another
and to other groups of the animal kingdom.

Fig. 252.—Diagrams of larvae. A, Loxosoma, × 208; a, anus; b, brain,

with left eye and ciliated pit; c, ciliated ring; ep, epistome; m,
mouth; o, oesophagus; st, stomach; x, aboral adhesive organ: B,
Cyphonautes larva of Membranipora (Electra) pilosa, × about 90;
a, m, o, st as in A; c, anterior part, and c', posterior part of the
ciliated ring; e, epidermis; ms, adductor muscle of shells; p,
pyriform organ, of unknown function; sh, shell; v, vestibule; the
"internal sac" or sucker, by which fixation is effected, is seen
between a and ms. (B, after Prouho.)
The comparative study of the larvae of the Polyzoa may be said to
date from 1877, when J. Barrois published an elaborate
Monograph[576] on this subject. Although some of Barrois' earlier
opinions have been subsequently modified, this work still gives the
best figures of the external form of the beautiful larvae of many
genera. A detailed account of the larval forms of Polyzoa must be
omitted from want of space; and the general conclusions only can be
given.

The larvae of the Entoprocta (Fig. 252, A) resemble the so-called

"Trochosphere" of Polychaeta (see p. 274). The common characters
shared by the larvae of Chaetopoda, Echiuroid Gephyrea, Mollusca,
and Polyzoa, and by adult Rotifera, may well point to the derivation
of these groups from a common ancestor. On this assumption, it is
possible that the Polyzoa have been derived from forms which
existed long ages ago, which combined the common characters of
these groups, and the structure of which we can picture to ourselves
only so far as the "Trochosphere" larva can be taken to represent it
in a much simplified condition. Such a view harmonises well with the
great antiquity of the Polyzoa. Certain Ectoproct forms have a larva,
known as Cyphonautes (Fig. 252, B), which closely resembles the
larval form of the Entoprocta; and it is a fact which probably has
considerable significance that this type of larva is known to occur
only in those species of Membranipora (Electra), Alcyonidium, and
Hypophorella, which lay eggs.[577] This may perhaps be regarded as
a primitive form of development which has been lost in species in
which development takes place inside the parent. Cyphonautes
compressus (Fig. 252, B), one of the commonest objects taken in the
surface-net off our own coasts, is the larva of Membranipora
(Electra) pilosa. Whilst this larva is provided with a well-developed
alimentary canal, those of most other Ectoprocta possess a mere
rudiment of this structure, and depend for their nutrition either on
yolk present in the egg or on material supplied by the parent. In most
cases the mature larva has no recognisable trace of a digestive
system; and, although it has a free-swimming period, it does not
become truly pelagic.
The alimentary canal of the larva of Pedicellina is known to persist in
the primary individual of the colony. In all other known cases, even in
that of Cyphonautes, the larva at fixation loses practically all its
internal organs, and becomes a mere body-wall containing a mass of
degenerated larval tissues. It is in fact a zooecium containing a
"brown body." A polypide-bud is now developed, the body-cavity
appears as the result of the shrinkage of the "brown body," and the
primary individual of the colony is thereby established.

The larvae of the Ectoprocta form a tolerably complete series,

starting from Cyphonautes, itself allied to the larva of the Entoprocta,
and ending with the Phylactolaemata. Alcyonidium (Fig. 253, B)
possesses a rudimentary alimentary canal,[578] although the most
conspicuous structures are those connected with the fixation and
other phenomena of larval life. The larvae of many of the encrusting
Cheilostomes (Fig. 253, A) resemble that of Alcyonidium, while those
of Bugula, Scrupocellaria, etc., belong to a type easily derivable from
that of the encrusting forms. The branching Ctenostomes
(Bowerbankia, etc.) have a larva which may be regarded as derived,
along slightly different lines, from that of Alcyonidium. The
Cyclostomata and the Phylactolaemata have the most modified
forms of larva. That of the former group may owe some of its
peculiarities to the occurrence of a remarkable process of embryonic
fission, which takes place in the ovicell, and as the result of which
each egg gives rise to a large number of larvae.[579] The
Phylactolaemata have a larva which is not unlike that of
Bowerbankia.
 Fig. 253.—A, Aboral view of free larva of Lepralia foliacea Ell. and Sol.;
a, long cilia of pyriform organ; g, aboral groove: B, longitudinal
section of embryo of Alcyonidium, × 135; c, ciliated ring; g, aboral
groove; m, mouth; n, nervous system; p, "pyriform organ," of
unknown function; s, "internal sac" or "sucker," by which fixation is
effected; st, stomach.

We have seen that the larva at fixation becomes a zooecium, which

in the Gymnolaemata forms a polypide-bud after fixation. The
peculiarities of the Phylactolaematous larva may be explained by
assuming that it becomes a zooecium while it is still free-swimming.
Thus the larva of Plumatella develops one or sometimes two
polypides, which actually reach maturity before fixation takes place.
That of Cristatella develops from two to twenty[580] polypides or
polypide-buds at the corresponding period, and it is in fact a young
colony while still free-swimming.

Now in most colonial animals, such as Coelenterates and Ascidians,

the larva metamorphoses itself into a temporarily solitary animal,
which then gives rise to the remainder of the colony by budding. The
majority of the Gymnolaemata behave in this way; while the
Phylactolaemata may not only develop a multiplicity of polypides in
their larval stage, but the individuality of the zooecia is then just as
much obscured as in the adult state. These facts are more easily
explained if we assume that Cristatella is the end-point in a series
than if we suppose it to be a starting-point.
On the view maintained by many authorities, that the Polyzoa are
related, through Phoronis, with the Gephyrea and the Brachiopoda,
we should expect to find in those Polyzoa which most closely
resemble Phoronis in their adult state—that is to say in the
Phylactolaemata—some indications of affinity to that animal in their
development. This is emphatically not the case. The hypothesis that
the Phylactolaemata are related to Phoronis leads, moreover, to the
improbable conclusion that the similarities between the Entoproct-
larva and Cyphonautes, on the one hand, and the Trochosphere
larva of Polychaeta, on the other hand, is entirely superficial and
meaningless. In spite, therefore, of the similarity between Phoronis
and a single individual of the Phylactolaemata, and in spite of the
marked resemblance between its nephridia and structures which
have been described in Cristatella[581] and Pectinatella[582] the
comparative study of the development appears to indicate that the
resemblances between Phoronis and the Phylactolaemata are the
result of a coincidence rather than of any close relationship.

A few points connected with the metamorphosis of the Polyzoa

deserve more special notice. There is generally great difficulty in
persuading larvae to fix themselves when kept in a small quantity of
water, which becomes over-heated in the air of a laboratory. The
difficulty may be surmounted by placing colonies containing
embryos, together with some clean pieces of the seaweed on which
the adults are habitually found, in a vessel closed by a piece of fine
muslin, and by leaving the vessel attached to a buoy or in a deep
tide-pool. The larvae being without an alimentary canal, fix
themselves, after a very short free life, on the seaweed.

It is probable that a great struggle for existence normally takes place

at the commencement of the metamorphosis. Any one who will
examine, in June or July, rocks covered by Fucus on which Flustrella
hispida is growing, will probably find numerous young fronds of
Fucus, from half an inch to an inch or two in length, growing under
the shelter of the older fronds. The bivalve larvae of Flustrella show
a marked preference for fixing on these young fronds—perhaps in
order that the duration of life of the colony may coincide with that of
the Fucus—and these young fronds are commonly covered by very
numerous recently-fixed larvae, and by young colonies of various
ages. Or, it is easy to observe, by placing pregnant colonies of
Bowerbankia in a vessel of water, that the larvae, which are hatched
out in thousands, fix themselves in dense masses on certain parts of
the wall of the vessel. It is clear that but a small proportion of these
larvae will find room for further development.

Next with regard to the mode of fixation. Attachment always takes

place by the surface on which the mouth or its rudiment is situated,
and the permanent alimentary canal opens on the opposite surface.
In Pedicellina, the one case in which the larval digestive organs are
known to become those of the first adult individual, this presupposes
a rotation of the alimentary canal, in order to bring it into its new
position.

It is well known that the larvae of other fixed animals may undergo a
somewhat similar change. Thus those of Ascidians and of Barnacles
fix themselves by their anterior end, and ultimately reach their adult
form by performing a kind of a somersault. The process may
perhaps be explained by supposing that some part of the anterior
end or of the oral surface is specially sensitive, and that the larva
fixes itself by that portion of its body which is best fitted for
ascertaining which is the proper substance on which to fix.

Budding.—The formation of a new individual may take place by the

outgrowth of part of the body-wall, as in Pedicellina (Fig. 243, p. 487)
and in Bowerbankia (Fig. 238, p. 480). In Pedicellina a young stalk is
formed by an outgrowth near one of the growing points, and the
upper part of this outgrowth becomes constricted off to form the
calyx. In other cases (cf. the growing ends of the branches in Fig.
237) a partition grows across the body-cavity at the growing edge of
the colony, and so cuts off a part destined to become a new
zooecium.
The zooecium formed in one of these ways acquires an alimentary
canal by the formation of a polypide-bud, some stages in the growth
of which are shown in Fig. 235 (p. 472). Contrary to what happens in
Coelenterates and Tunicates, in which the endoderm takes part in
the budding, there is good reason for believing that in Polyzoa the
polypide-bud is developed entirely from ectoderm and mesoderm.
[583] The bud is a two-layered vesicle, attached to the inner side of
the body-wall. Its inner layer is derived from the ectoderm, which at
first projects into the body-cavity in the form of a solid knob
surrounded by mesoderm-cells. A cavity appears in the inner,
ectodermic mass, and the upper part of the vesicle so developed
becomes excessively thin, forming the tentacle-sheath, which is
always developed in the condition of retraction. The lower part
becomes thicker; its inner layer gives rise to the lining of the
alimentary canal, to the nervous system, and to the outer epithelium
of the tentacles, which grow out into the tentacle-sheath (cf. Fig.
235). The outer layer gives rise to the mesodermic structures, such
as the muscles, connective tissue, and generative organs.

These processes are fundamentally similar, whether in the

metamorphosed larva, in a young zooecium, in an old zooecium
after the formation of a "brown body," or in the germinating statoblast
of the Phylactolaemata.

CHAPTER XIX

POLYZOA (continued)

CLASSIFICATION—GEOGRAPHICAL DISTRIBUTION—PALAEONTOLOGY—
METHODS FOR THE EXAMINATION OF SPECIFIC CHARACTERS—
TERMINOLOGY—KEY FOR THE DETERMINATION OF THE GENERA OF
BRITISH MARINE POLYZOA
Our account of the Polyzoa would be manifestly incomplete without
some reference to the systematic arrangement of these animals. An
outline of the principal groups has been given on p. 475. So far, the
classification is easy, but it is otherwise when we attempt to
subdivide most of the groups any further.

Systems of classification which depend exclusively upon the external

characters of animals have been repeatedly shown to be
unsatisfactory. Now with regard to the Polyzoa, not only is it the case
that the great majority of forms are only known in their external
characteristics, but current systems of classification cannot be
regarded as final, because it is not yet certain which of the external
features have most systematic value. Two obvious points can be at
once selected—namely, the character of the zooecium and the
character of the entire colony. One or two instances will serve to
show what different results are obtained by depending exclusively on
either of these characters by itself.

According to the older writers, the habit of the colony was taken as
the most important generic character; and there can indeed be no
doubt that this feature has great importance within certain limits. Any
one who has examined different species of such genera as Flustra,
Cellaria, Bugula, Retepora, etc., must feel that the form of the colony
goes for a good deal. But a consideration of other cases shows that
there is great risk in the indiscriminate use of this method of
arranging the Polyzoa. The old genus Eschara, composed of forms
with an erect coral-like habit,[584] included species which are now
placed in such different genera as Lepralia, Porella, Microporella,
etc. The older works on Polyzoa include all encrusting forms of
Cheilostomata, with a completely calcareous front wall, in the genus
Lepralia, the members of which are now distributed in numerous
widely separated genera.

As an instance of the converse arrangement—essential similarity of

the zooecia with great differences of the general habit—may be
mentioned the common Membranipora (Electra) pilosa.[585]
Ordinarily growing in the form of close encrustations on seaweeds,
this species may take on entirely different habits of growth. The
zooecia are now dissociated, growing in single lines over the
substratum; now forming erect tufts, composed of single lines of
zooecia or of several rows. The erect, branching habit appears to be
induced in the first instance by the character of the seaweed on
which the colony begins life. Thus colonies which encrust the thin
branches of Corallina may have impressed on them something of the
mode of growth of the seaweed, so that when they extend beyond
the tips of the branches of the Corallina, they continue to grow in
delicate branches, which still retain more or less the same diameter
as those which form their base. An extreme variation results in the
beautiful form known as Electra verticillata, in which the zooecia are
arranged with great regularity in whorls, which together form erect
branches.[586] But with all these variations, the zooecia are so much
alike that it is hardly possible to regard the extreme forms as more
than varieties of a single species. A careful examination of this case
would convince most observers that the characters of the zooecium
are a more trustworthy guide to classification than those of the entire
colony, a result which was first clearly stated by Smitt, and amply
confirmed by Hincks.[587]

The avicularia of the Cheilostomata afford useful help in classifying

this group; but while certain genera are always provided with
avicularia, others include some species with these organs, and other
species without them. Again, while the species of some genera (e.g.
Cellepora) possess a great variety of forms of avicularia, the same
pattern of avicularium may characterise several widely different
genera. Further, the position of the avicularium may be very different
in species which are apparently closely related. Well-developed
vibracula, although constant in their occurrence in such forms as
Scrupocellaria (Fig. 254) and Caberea (Fig. 242), occur here and
there in species of encrusting forms which are ordinarily placed in
very different families.
Now although some of these discrepancies are perhaps due to
errors in classification, whereby species which are really allied have
been wrongly placed in distinct genera, this explanation would not
prove satisfactory in all cases. Thus in Bugula, a genus which is
specially characterised by the high development of its avicularia,
these organs are normally absent in B. neritina. The fact that this
species was rightly placed in the genus has been confirmed by the
discovery made by Waters[588] that avicularia occur in specimens
which are believed to be identical with that species.

Fig. 254.—A, Front view, and B, back view of part of a branch of

Scrupocellaria scabra, Van Ben., Durham Coast, × 43; a, lateral
avicularium; a', smaller median avicularium; ap, membranous
aperture; f, fornix; r, rootlet; s, seta of vibraculum; v.z, vibracular
zooecium.

1. The Cyclostomata appear to fall naturally into two main groups,

(A) the Articulata, including the Crisiidae (Fig. 237), distinguished
by their erect branches, divided at intervals by chitinous joints; and
(B) the Inarticulata, which include the remaining families, whether
erect or encrusting, agreeing in the negative character of being
unjointed.

2. The Cheilostomata consist of (A) the Cellularina, including the

flexible, erect forms, such as Bugula (Fig. 233) and Scrupocellaria
(Fig. 254); (B) the Flustrina, to which belong Flustra (Fig. 232),
Membranipora (Fig. 256, A, B), Micropora (Fig. 256, C), and other
forms in which the front wall of the zooecium is either membranous,
or depressed and marked off by a ridge-like margin; (C) the
Escharina, including the great majority of forms, in which no part of
the front wall remains membranous, the wall of the zooecium being
wholly calcified.

3. The Ctenostomata comprise (A) the Alcyonellea or encrusting

forms; and (B) the Vesicularina or branching forms. The zooecia in
the latter subdivision (Fig. 238) are given off from a tubular stem or
stolon, which is usually erect and branching.

We thus have the following arrangement of recent forms. The genera

mentioned are for the most part those which have already been
alluded to in the preceding account:—

Sub-class I. Entoprocta.
Loxosoma, Pedicellina, Urnatella.

Sub-class II. Ectoprocta.

Order 1. Gymnolaemata.
Sub-order 1. Cyclostomata.
A. Articulata. Crisia.
B. Inarticulata. Hornera, Idmonea, Tubulipora,
Stomatopora,
Diastopora, Entalophora, Lichenopora.
Sub-order 2. Cheilostomata.
A. Cellularina. Aetea, Eucratea,[589] Catenicella,
Cellularia, Gemellaria, Menipea, Scrupocellaria,
Caberea, Notamia (= Epistomia), Bicellaria, Bugula,
Beania.
B. Flustrina. Cellaria, Flustra, Membranipora, Electra,
Lunulites, Membraniporella, Cribrilina, Micropora,
Selenaria.
C. Escharina. Retepora, Microporella, Lepralia, Porella,
Smittia, Mucronella, Schizoporella, Schizotheca,
Mastigophora, Porina, Cellepora.
Sub-order 3. Ctenostomata.
A. Alcyonellea. Alcyonidium, Flustrella.
B. Vesicularina. Vesicularia, Amathia, Bowerbankia,
 Farrella, Hypophorella, Triticella, Mimosella,
Victorella, Paludicella.
Order 2. Phylactolaemata.
Fredericella, Plumatella (including Alcyonella), Lophopus,
Cristatella, Pectinatella.

Even this classification, which deals only with the larger groups,
must not be made use of without a word of warning. The division of
the Cheilostomata is a matter of great difficulty; and no scheme
which has yet been suggested can be regarded as more than
tentative. The great number of forms included in this group makes its
subdivision extremely desirable from the point of view of
convenience; but a further knowledge of the anatomy and of the
development of many of the forms of doubtful systematic position is
probably necessary before any scheme which is likely to be
permanent is put forward. Those who desire to make a further study
of the classification of the Polyzoa should refer to the works of
Hincks,[590] Busk,[591] MacGillivray,[592] and Gregory.[593]

The Polyzoa do not appear to lend any valuable assistance towards

settling the disputed problems of Geographical Distribution. They are
not in any case terrestrial, while the fresh-water species do not
always respect the limits between the great zoogeographical
regions. It has already been pointed out (p. 504) that Plumatella,
Fredericella, and Lophopus are believed to occur in Australia, and
the first-named genus is practically world-wide in its distribution.

Many marine forms also have a surprisingly wide distribution. Thus

among the British species which are described by Mr. Hincks as
occurring from Norway to New Zealand are Membranipora pilosa,
Scrupocellaria scruposa, Cellaria fistulosa, Microporella ciliata, and
M. malusii. Even if it should be proved that specific differences do
exist between the southern forms and our own, there can be no
doubt of the wide distribution of certain species. It was pointed out by
D'Orbigny that Bugula neritina has the habit of attaching itself to the
bottoms of ships, a fact which may possibly account for the wide
distribution of this species; although it would not be safe to assume
this explanation of the facts in all cases. Other Polyzoa, on the
contrary, have a more restricted range. Thus Catenicella is specially
characteristic of the Australian region.

It is perhaps surprising that marine Polyzoa should in so many cases

have so wide a range. Even though it is the rule for Polyzoa to have
free larvae, the period during which these larvae are free-swimming
is, so far as is known, a short one in most cases. Cyphonautes is a
common pelagic form (see p. 510), and probably remains for a
considerable period in the larval condition. Other Polyzoon-larvae
appear to fix themselves very soon after their birth; and this would
not appear to give much time for them to be carried to great
distances by ocean-currents. It may, however, be suggested that it
does not follow that because we know that a larva may, under
favourable conditions fix itself a few minutes after it becomes free,
we should be justified in assuming that that larva would not retain for
a long period the power of undergoing a normal metamorphosis
should it be drifted away from suitable fixing-grounds.

Palaeontology.[594]—The number of fossil Polyzoa is enormous.

D'Orbigny devoted two hundred plates and more than a thousand
octavo pages[595] to a Monograph on the Cretaceous Polyzoa of
France. Many of the fossil forms are extraordinarily well preserved,
and there is often no difficulty in recognising the identity between
certain fossil species belonging to the more recent formations and
living forms. It thus becomes necessary to consult Palaeontological
memoirs in working at recent Polyzoa.

While the great majority of fossil Polyzoa do not differ in any

essential particular from recent species, this is not altogether the
case with the Palaeozoic forms. Leaving out of account the
Stromatoporoids, which have been variously referred to the
Sponges, Hydrozoa, and Foraminifera, as well as to the Polyzoa, the
Palaeozoic strata contain large numbers of peculiar Cyclostomata,
together with members of the Trepostomata, a fourth Sub-order of
Gymnolaemata, allied to the Cyclostomata. The Trepostomata are
for the most part Palaeozoic, but a few survived as late as the
Jurassic period.[596] These, with the other Polyzoa from the same
formations, are considered by Dr. Gregory in his recently published
Catalogue of the Fossil Bryozoa in the British Museum (1896).

The number of Polyzoa recorded from the earlier secondary strata is

small. The majority of the known Jurassic forms belong to the
Cyclostomata; and one or two Cheilostomes are recorded from the
same period. Recent papers by Walford[597] on Jurassic Polyzoa
contain the description of genera which are believed to be
intermediate between the Cyclostomata and Cheilostomata,
particularly with regard to the characters of their ovicells. Although it
is not impossible there may be a connection between the ovicells of
these two groups, it has yet to be proved that the two sets of
structures are homologous.

The Cretaceous period marks the commencement of a large number

of Cheilostome genera, although the Cyclostomes still remain
numerous.

In the Tertiary formations the Cyclostomes gradually become less

numerous, and although in earlier geological periods they far
outnumbered the Cheilostomes, these relations are now reversed.
Certain Tertiary strata, and particularly the Coralline Crag (Pliocene),
are remarkable for the extremely large number of Polyzoa they
contain. It will be noticed that no mention has been made of the
Entoprocta, the Ctenostomata, and the Phylactolaemata. Their
absence in the fossil condition[598] need not, however, be a matter
for surprise, as none of these forms are so well suited for being
fossilised as are the calcareous Cyclostomata and Cheilostomata.
There is consequently no adequate reason for assuming that the
absence of a palaeontological record implies that these groups have
been recently evolved.
Determination of Genera of Marine Polyzoa.—The species to
which a Polyzoon belongs can only be determined, in most cases,
with the assistance of the low powers of a microscope. There are
very great advantages in the use of a binocular instrument, by
means of which a microscopic preparation appears with its parts
standing up in proper relief.

In the case of the calcareous forms, the external characters may be

more readily made out in a dry preparation than in any other way.
For this purpose, the colony should be washed with fresh water, in
order to remove the salts, which otherwise crystallise out on drying
and obscure the surface. Preparations of this kind must be looked at
with the aid of reflected light. Canada-balsam or glycerine
preparations are also valuable, whether stained or unstained; and
are essential for the examination of the softer forms. In the case of
erect species, both surfaces of the branch should be looked at. The
opercula, avicularia, and rosette-plates afford important systematic
characters in the case of the Cheilostomata.

It must not be forgotten to take account of the condition of the

zooecia at different ages. The old zooecia often become entirely
altered in form, by the deposition of additional calcareous matter, or
by the loss of certain parts present in the younger zooecia. Thus the
marginal spines may be entirely lost in the older individuals, while in
those forms which develop a "peristome" (see Fig. 255 and p. 524),
the characters of the orifice can often be determined in the young
zooecia only. It is thus essential to examine the growing ends of the
branches or the rim of the colony, as the case may be.

Fig. 255.—Illustrating the nature of a secondary orifice

(Cheilostomata). A, Mucronella coccinea Abildg., Scilly Is., × 40.
 The ovicell (o) overhangs the primary orifice, which is concealed
by the great development of the peristome, produced into the
mucro (mu); t, the three teeth (denticles) within the secondary
orifice; a, avicularium. B, Porella compressa Sowb., Norway, × 40;
p.o. primary orifice, above which is a concave lamina, the
beginning of the ovicell. In the lower zooecium the ovicell (o) is
further grown. The primary orifice is still visible, but it is partially
concealed by the growth of the peristome, which encloses a
minute avicularium; m, mandible of avicularium. C, Older part of
the same colony; pr, peristome; s.o, secondary orifice; o', adult
ovicell; p, pores.

In order to make preparations with the tentacles expanded,

hydrochlorate of cocaine, chloral hydrate or spirit should be added
gradually to the water. When the animals are completely
anaesthetised they may be killed by means of a 7-10 p.c. solution of
sulphate of copper (best made in distilled water or in rain water).
This method gives admirable results in the case of both fresh-water
and marine Polyzoa. The use of formaline (see p. 229) may be
strongly recommended for the Vesicularina.

The only recent work dealing with all the marine British forms is Mr.
Hincks' invaluable History of the British Marine Polyzoa.[599] As the
use of this book, unaided by any artificial help, is by no means easy
to the beginner, the following key has been compiled as an index to
the genera. The Entoproct forms, Loxosoma and Pedicellina (see pp.
488-491), are not included in the table.

Fig. 256.—Illustrating the terminology of the front surface of the

zooecium (Cheilostomata). A, Membranipora (Electra) pilosa L.,
Cromer, × 47; ap, the membranous "aperture;" o, orifice. B,
Membranipora flemingii Busk, Plymouth, × 60; ap, the aperture,
 enclosed in a calcareous "area" (a); av, avicularium; s, marginal
spines. C, Micropora coriacea Esper, Plymouth, × 43; a, area
(calcareous); o, operculum; ov, ovicell.

In order to facilitate the use of the table here given in conjunction

with Mr. Hincks' work, the nomenclature there adopted has been
followed throughout. References to other descriptions of the species
may be obtained by consulting Miss Jelly's admirable Synonymic
Catalogue of the Recent Marine Bryozoa.[600]

Terminology.—A few technical terms must of necessity be

employed. The colony is adherent when its zooecia are attached to
the object on which the colony is growing. The zooecium is the body-
wall of a single individual; and, except in transparent species, is the
only part which can be seen from the outside in the retracted
condition of the polypide or tentacles with the alimentary canal. The
outermost layer of the zooecium is known as the ectocyst; it may be
simply membranous, or calcified, or may be rendered opaque by
foreign bodies; its surface in calcareous forms is often marked by
pores (Fig. 239, C, p), which are vacuities in the calcareous wall,
closed externally by membrane. A special median pore (Fig. 241, A,
m.p) may occur, and is in some cases at least a complete perforation
through the body-wall.

The tentacles are protruded through the orifice, which in

Cheilostomata is usually guarded by a movable chitinous lid, or
operculum (Fig. 256, A, o). Should the ectocyst be thickened or
raised into a ridge surrounding the orifice, a tubular passage results,
known as the secondary orifice (Fig. 255), at the deeper end of
which is the true orifice. The peristome (Fig. 255, C, pr) is the raised
or thickened part which gives rise to the secondary orifice. Should
the zooecium be outlined by a raised ridge, the part so enclosed is
known as the area (Fig. 256, C, a), if calcareous. The aperture or
opesia (Fig. 256, A, B, ap) is a membranous part of the front surface;
and may consist of the whole or part of the area. The orifice or the
aperture is commonly provided with spines (Fig. 256, B, s).
 Fig. 257.—A, Cribrilina annulata Fabr., Norway, × 33; c, calcareous
bars concealing the membranous aperture: B, Membraniporella
nitida Johnst., Plymouth, × 45; a, calcareous bars growing up
round the margin of the aperture; b, the same, further developed;
c, the same, completely formed (as in A); av, avicularium; o,
immature, and o', mature, ovicell; s, marginal spines.

The avicularium and the vibraculum are specially modified zooecia

(see p. 482), which occur in a great variety of forms, in certain
Cheilostomata only. The operculum of the ordinary zooecium is
represented by the mandible (Fig. 239, B, m) in the avicularium, and
by the seta (Fig. 242, s) in the vibraculum. The representative of the
zooecium itself is known as the avicularian (Fig. 239, A, a.z) or
vibracular zooecium (Fig. 242, v.z).

An ovicell is a swelling in which the embryo develops, in certain

Cyclostomata (Fig. 237) and Cheilostomata (Fig. 241, A, o). A stolon
(Fig. 238, B, st) is a stem, not formed of fused zooecia, from which
new individuals originate. An internode, in a jointed colony, is the
part between any two joints. The fornix or scutum (Fig. 254, A, f) is a
modified spine which in some Cheilostomata overhangs the
aperture. A mucro (Fig. 255, A, mu) is a spike or protuberance
developed just below the orifice. A sinus (Fig. 239, B, s) is a slight
bay on the lower margin of the orifice.

The orifice opens at the upper end of the zooecium, on its front
surface. The length of the zooecium is the distance from the upper to
the lower ends, and the width the distance between its sides.

1. One or more of the following characters: orifice provided with

 an operculum; avicularia or vibracula present; a globular
ovicell above the orifice of certain zooecia (Cheilostomata)
7
Opercula, avicularia, vibracula, and ovicells completely
absent, or inconspicuous. Calcareous or non-calcareous. If
calcareous, the orifice is not at the end of a free cylindrical
portion
3
Calcareous; zooecia cylindrical, often united for the greater
part of their length, but usually ending in a free cylindrical
portion, which bears the terminal orifice. The zooecia may
be much obscured by calcifications surrounding their basal
parts
2

2. Zooecia long, tubular, with a lateral membranous region at

the upper end, given off quite separately from a creeping
stolon
Aetea
Zooecia more or less united to one another, orifice without
chitinous operculum (Cyclostomata[601])
63

3. Zooecia without marginal spines; arising from a branching

axis, which is not formed of zooecia
74
Colony adherent; or erect, fleshy and slightly branched; or
erect, encrusted with earthy matter and repeatedly branched
72
Characters not as above 4

4. Zooecia minute, boat-shaped, united by a delicate tube.

Aperture large, with marginal spines
Beania mirabilis
Colony delicate, erect; zooecia wider above than below 5

5. No marginal spines 6