EMBRYOLOGY

Development of lymph node(26):

Lymph nodes are encapsulated centers of lymphocyte differentiation and

proliferation,belonging to the class of secondary or peripheral lymphoid tissue.

A normal young adult body contains about 400-450 lymph nodes. The head and

neck carry some 60-70 nodes;arm and superficial thoraco-abdominal wall upto umbilicus,

about 30 nodes; the leg, infraumbilical abdominal wall and perineum about 20 nodes. The

remaining is divided between thorax about 100 nodes, and abdomen and pelvis about 230

nodes. Most richly served by nodes are gastrointestinal tract and tracheo

bronchopulmonary tract.

The first indication in the embryo of the establishment of lymphatic

system becomes evident in the 6th week of development.When the lymphatic vascular

channels have been fairly well sketched out,lymph nodes begin to make their appearance

in the system. Certain areas of connective tissue are colonized by lymphoblasts and

lymphocytes are formed in large numbers in loose mesh of the young connective tissue.

Such aggregation of lymphocyte inserted on the course of a lymphocytic channel break it

up into meshwork of tortures, smaller channels interlacing through the lymphoid tissue.

At the same time small blood vessels follow along the connective tissue frame work, in

which lymphocytes proliferate, gradually, as the node takes shape cords of dense

lymphoid tissue develop along the vascular strands of connective tissue.

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 These cords of lymphoid tissue establish the characteristic picture one sees

in the medulla of a lymph node. At a relatively later stage of development.

Massive spheroidal aggregations of lymph are formed in the zone of the

developing node, establishing its cortex; the extensions of the growing mass of

lymphoid tissue compress the immediately shrouds connective tissue to join the

capsule

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 ANATOMY:

Lymph node structure 27:

Lymph nodes are small oval or reniform bodies 0.1-2.5 cm long lying in the

course of lymphatic vessel. Each usually has a slight indentation on one side,the ‘hilum’,

through which blood vessels enter and leave .

Lymph nodes have a highly cellular cortex and a medulla, containing numerous,

poorly demarcated cavities. The cortex is deficient at the hilum, where the

medulla reaches the surface; thus the efferent vessel emerges from the medulla,

while afferent vessels empty into the cortex5.

Topographical naming of lymphnodes:

Cervical lymphnodes are grouped in to :

 superficial circle of cervical lymphnodes

 Deep cervical lymphnodes

Deep cervical lymph nodes for convenience of description are often grouped as :

 Anterosuperior 􀃆 includes jugulo diagastric nodes

 Anteroinferior

 Posterosuperior

 Postero inferior 􀃆 Includes jugulo omohyoid nodes

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Superficial circle of cervical lymph nodes is made up of following groups.

 Sub mental

 Sub mandibular

 Buccal and mandibular (facial)

 Preauricular (parotid)

 Post Auricular (mastoid)

 Occipital

 Anterior Cervical nodes

Axillary lymph nodes are divided into five groups .

 Anterior or pectoral group

 Posterior or scapular group

 Lateral group along the upper part of humerous

 Central group

 Apical or infraclavicular group.

Other lymph nodes draining upper limb include :

 Infra clavicular nodes

 Deltopectoral nodes

 Superficial cubital or supratrochlear nodes

 Deep cubital lymph nodes at the bifurgation of the brachial artery

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Lymph nodes of posterior abdominal wall include :

 External iliac

 Common iliac

 Lumbar / aortic group – It is further divided into

o Preaortic nodes - Coeliac

 Superior mesenteric

 Inferior mesenteric

o Lateral aortic nodes

o Retroaortic nodes

Lymph nodes draining lower limb include :

 Superficial inguinal nodes consisting of

o Lower vertical group

o Upper lateral group

o Upper medial group

 Deep inguinal nodes – They lie medial to the upper part of femoral vein.

The most proximal nodes of this group; gland of Cloquet or of

Rosenmullar, lies in the femoral canal.

 Popliteal lymph nodes – These lie near the termination of small saphenous

vein, deep to deep fascia.

 Anterior tibial node.

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HISTOLOGICAL ORGANISATION (28)

A lymph node consists of an encapsulated mass of lymphoid tissue

traversed by specialized lymph vessels called lymphatic sinuses. Its connective tissue

framework consist of a capsule which invests the whole organ but which is greatly

thickened at the hilus.

The efferent lymphatics are continues with a sub-capsular sinus, from there

the lymph flow inward through narrow cortical sinus and then to medullary sinus that

carry the lymph to the efferent lymphatic draining the node.

Lymph sinus:

The afferent lymphatic vessel approach the convex surface of the node traverse in

capsule obliquely and open into the marginal or sub capsular sinus.

Examined under the scanning electron microscope the sinus appear as channels

lined by a layer of attenuated squamous cells with the lumen bridged by a mesh work of

satellate reticular cells connected to each other and a wall of the sinus via slender cells

processor.

At present it is widely believed that there are two distinct categories of cells lining

the sinuses 1) Microphages and 2) flattened satellite organ of blood and lymph vessels,

are inconspicuous and they take up only small amount of particulate matter by

endocytosis.

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Cortex:

The cortical parenchyma appears with the high microscope, as a dense mass of

lymphoid cells traversed in places by the collageneous trabecular and intermediate

sinuses. Certain regional differentiation with in the cortical parenchyma have been

traditionally been classified as primary lymphoid nodule which are spherical or ovoid

areas of tightly packed small lymphocyte that are discernible, within the continum of

lymphoid tissue comprising the cortex.

Secondary nodules have a paler central zone, called the germinal center made up

of large lymphocyte and macrophage. The internodular cortex and inner or deep cortex

consist of different lymphoid tissue devoid of germinal center.

There is no distinct boundary between the outer and inner cortices and the latter

continues into the medullary cord without any discernible demarcation.

Medulla:

The medullary cord consists of aggregations of lymphoid tissue organized around

small blood vessels.They are made up of small lymphocyte, plasma cell and macrophage

occupying the in the interstice of a rich network of reticular fiber and associated reticular

cells. The parenchyma of the lymph nodes may normally contain small number of

granulocytes and they may be greatly increased on stimulation or in pathological

conditions.

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The paracortex:

It contains small T-type lymphocytes that are suspended in nests composed of

interdigitating reticulum cells. Under antigenic stimulation, the T-type lymphocytes may

evolve into T-type immunoblasts.

Cells (22) :

Most of the cells in the lymph nodes are lymphocytes, both B- lymphocytes and

T lymphocytes are present.

The distribution of lymphocytes varies in different regions. In the cortex cells are

densely packed and form isolated lymphoid follicles that are composed mainly of B-

lymphocyte.As a consequence of antigenic stimulation, the small B-lymphocytes within

the germinal centers are transformed into large cells that have large, round nuclei and

prominent nucleoli. Previously known as reticulum cells, these large cells have been

designated by Lukes and Collins (1974) as large noncleaved cells and by Lennert (1967)

as centroblasts.

The transformation of the small lymphocytes into the large noncleaved cells

proceeds through several stages and forms, identified by Lukes and Collins (1974) as

small cleaved cells, large cleaved cells, and small noncleaved cells. The large noncleaved

cells may participate in further lymphopoiesis or transform into immunoblasts which,

outside of the germinal centers, evolve into plasma cells. Cumulatively, the transformed

lymphocytes may be designated as follicle center cells.

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 B-lymphocytes mature to plasma cells and are seen in the medullary cords. T-

lymphocytes are found primarily in the area between the follicles and in the deep cortex

(known as Para cortex).

Macrophages also occur and are present mainly in the walls of sinuses, medulla

and few within germinal centers. Other cells present are fibroblasts, endothelial cells,

granulocytes,pericytes, smooth myocytes, follicular dendritic cells and myelinated nerve

fibers.

Under antigenic stimulation,the whole node reacts by increase in size and

vascularity due to proliferation of cells like lymphocytes and macrophages, the number

and size of germinal centers also increase.(22)

Normal Cytology :

The lymphocytes constitute 87%-99%, the plasma cells 0-5%, and remainder cells

form 1%-3% of the total population in the aspirate from a normal lymphnode.28

1. Mature lymphocytes

The small, round lymphocyte measures 7- 10 μm in MGG (may grunwald

giemsa) stained smears. It has thin rim of pale cytoplasm often visible at one edge of the

cell and nuclei characterized by blocks or clumps of dense chromatin.28

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2. Follicular center cells

Follicular center cells are of following types :

(a) Small cleaved lymphocytes : These are 10-12μm in diameter, their nuclei show deep

indentation of nuclear membrane, and hence the term cleaved and is surrounded by a

moderate amount of cytoplasm.28

(b) Small non-cleaved lymphocytes : These are larger measuring 12-20μm in diameter,

they are round and have a scanty amount of cytoplasm unlike mature lymphocytes and

their non-cleaved nuclei contain prominent nucleoli.28

(c) Large cleaved lymphocytes: These are round cells 20-40μm in diameter and nuclei

appear cleaved as a result of a deep indentation of the nuclear membrane and nucleoli are

not prominent and the cells have abundant basophilic cytoplasm.28

(d) Large non-cleaved lymphocytes: These are 20-40μm in diameter. A moderate

amount of cytoplasm surrounds a large round vesicular nucleus. The nucleus has smooth

outline and contain two or three peripheral nucleoli.28

3. Immunoblast

These are large cells measuring 20-40μm in diameter, three times larger than

small lymphocytes. The nuclei are round with irregular, finely granular chromatin and

one or more eccentrically placed nucleoli, and the surrounding cytoplasm is abundant

with cytoplasmic vacuolization.28

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4. Plasma cells

In plasma cells the nuclei are eccentrically placed and possess densely packed

coarse chromatin that may be arranged in a typical cart-wheel like pattern and the

cytoplasm is deeply basophilic and contains a para-nuclear clear zone.28

5. Macrophages

These cells show wide variation in size, in the resting phase the cells measure 14-

34μm in diameter. Those containing phagocytosed fragments of degenerated cells

(tingable bodies) may measure upto 50μm. The nucleus is about 13μm in diameter and

contains evenly distributed reticulated chromatin, one to three small nucleoli may be

present and at times such macrophages possess two or more nuclei.28

Other cells like interdigitating reticulum cells, endothelial cells can also be seen in

normal lymphnode aspirate. Eosinophils, plasma cells and neutrophils may also be seen

depending on the pathologic process.28

6. Lymphoid globules (lymphoglandular bodies)

They are the rounded cytoplasmic fragments measuring up to 8μm in diameter

and are scattered in the background. The fragments stain an even pale blue identical to

the cytoplasm of intact cells with Giemsa stain. They are characteristic finding in smears

of lymphoid tissue. They differ from necrotic debris by their regular round shape and

their uniform staining..

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 FUNCTIONS OF LYMPH NODE

1. It provides the provision of a labyrinth of channels of large volume and surface

area,through which lymph slowly percolates.

2. The exposure of foreign material in the lymph to macrophages in nodal sinuses.

3. The trapping of antigens by different phagocytes.

4. Production of lymphocytes and a pool of stem cells able to become antibody

producing, B-lymphocytes and mature T-lymphocytes.

5. Humoral antibody production.

6. When lymph passes through the lymph nodes it is filtered. The water and

electrolytes are removed, proteins and lipids are retained.

7. Lymph flow plays important role in redistribution of fluid in the body.

Lymph nodes are highly effective filter interposed in path of lymph drainage form

nearly all region of the body. One of their functions is to limit the dissemination of

bacteria and malignant cell by their removal from the lymph before it reaches the blood

via the thoracic duct.When there is an infection in a distant part of the body, the regional

lymph node become inflamed as a result of the localization of bacteria or toxin carried in

the lymph to the gland.

The lymph node contains a very efficient filtration system in the cortical and

medullary sinus, these are lined with phagocytic cell that engulf bacteria and red cells or

other particulate material. The efficiency of the filtration system can be demonstrated by

the direct perfusion of pathogenic bacteria into the lymphatic that are afferent to a lymph

node, simultaneous culturing of the efferentlymphatic will show to be sterile. However

lymph node are much less efficient barrier to lymph borne cancer cell.(29)

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