1. Amino acids.  2014 august 29.

bp8, a novel peptide from avian immune system, modulates b cell

developments.
Author: Liu XD, Zhou B, Feng XL, Cao RB, Chen PY.

Division of Key Lab of Animal Disease Diagnosis and Immunology of China's

Department of Agriculture, Nanjing Agricultural University, Nanjing, People’s
Republic of China.

Abstract
The bursa of Fabricius (BF) is the key humoral immune organ unique to birds, and
is critical for early B-lymphocyte proliferation and differentiation. However, the
molecular basis and mechanisms through which the BF regulates B cell
development are not fully understood. In this study, we isolated and identified a
new bursal peptide (BP8, AGHTKKAP) by RP-HPLC and MALDI-TOF-MS. BP8
promoted colony-forming pre-B formation, bound B cell precursor, regulated B cell
development in vitro as well as in vivo, upstream of the EBF-E2A-Pax5 regulatory
complex and increased immunoglobulin secretion. These data revealed a bursal-
derived multifunctional factor BP8 as a novel biomaterial which is essential for the
development of the immune system. This study elucidates further the mechanisms
involved in humoral immune system and has implications in treating human
diseases.

 2. Peptides. 2012 May; Epub 2012 March 10.

Isolation, modulatory functions on murine B cell development and
antigen-specific immune responses of BP11, a novel peptide from
the chicken bursa of Fabricius.

Author information : Liu XD1, Feng XL, Zhou B, Cao RB, Li XF, Ma ZY, Chen PY

Division of Key Lab of Animal Disease Diagnosis and Immunology of China's

Department of Agriculture, Nanjing Agricultural University, Nanjing, China .

Abstract
The bursa of Fabricius (BF) is the central humoral immune organ unique
to birds which plays important roles in B lymphocyte differentiation. Here, a new
bursal peptide (BP11) with the amino acid sequence DVAGKLPDNRT was
identified and characterized from BF. It was proved that BP11 promoted CFU pre-B
formation, and regulated B cell differentiation, including increase the percentage of
immature and mature B cells in BM cells co-cultured with IL-7. BP11 also exerted
immunomodulatory function on antigen-specific immune responses in BALB/c mice
immunized with inactivated influence virus (AIV, H9N2 subtype) vaccine, including
enhancing AIV-specific antibody and cytokine production. Furthermore, it was
noteworthy that BP11 stimulated antibody productions and potentiates the Th1 and
Th2-type immune responses in dose-dependent manner in chicken. These results
suggested that BP11 might be highly relevant for the development of avian
immune system.

3. Regul Pept. 2013. Epub 2013 July 25.

Comparison of immunomodulatory functions of three peptides
from the chicken bursa of Fabricius.

Author information: Liu XD1, Feng XL, Zhou B, Cao RB, Li XF, Ma ZY, Chen PY

Division of Key Lab of Animal Disease Diagnosis and Immunology of China's

Department of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.

Abstract
The bursa of Fabricius (BF) is the acknowledged central humoral immune organ
unique to birds which plays important roles in B cell development and antibody
production. Little information on immunomodulatory functions of BF is reported,
except for several reported active bursal-derived peptides. Three peptides were
identified and characterized from BF through RP-HPLC and MADIL-TOF methods.
They are named as bursal peptide (BP)-I, BP-II, BP-III. These peptides promoted
CFU pre-B formation and decreased PU.1 expression. The different
immunomodulatory activities of these three bursal peptides on antibody and
cytokine productions were verified by the immunization comparative experiment.
The results showed the three bursal peptides enhanced AIV-specific antibody and
cytokine production, T-cell immunophenotyping at reachable concentrations. These
results indicate the important orientations for the comprehensive understanding of
the humoral central immune system, and provide a novel insight on new
experimental reagents for immuno-adjuvant or immunopharmacological.

4. Peptides. 2012. Epub 2012 January 24.

Characterization and immunomodulatory function comparison of
various bursal-derived peptides isolated from the humoral central
immune organ.
Author information: Feng XL, Liu QT, Cao RB, Zhou B, Zhang YP, Liu K, Liu XD,
Wei JC, Li XF, Chen PY.

Division of Key Lab of Animal Disease Diagnosis and Immunology of China's

Department of Agriculture, Nanjing Agriculture University, Nanjing 210095, China.

Abstract
The bursa of Fabricius (BF) is the acknowledged central immune organ, which is
important to the B cell differentiation and antibody production. However, due to
difficult purification, the immunomodulatory peptides from BF were little reported. In
this study, the extract samples of BF were taken to a chromatographic analysis by
RP-HPLC. Five novel low molecular weight peptides were isolated from BF, with
amino acid sequences of YEYAY, RMYEE, GPPAT, AGCCNG, and RRL, and
named as Bursal pentapeptide (BPP)-III, -IV, -V, and Bursal hexapeptide (BHP),
and Bursal tripeptide (BTP), respectively. BSP-I, BSP-II, BPP-I and BPP-II are
recently reported to be the bursal-derived bioactive peptides. In this paper, we
analyzed the chemical formula and characteristics of these nine bursal-derived
peptides. The immunization comparative experiment verified the different
immunomodulatory activity of these nine bursal peptides on antibody and cytokine
productions. Furthermore, the results showed that at reachable concentrations,
BPP-II and BPP-I induced antibody productions, lymphocyte viabilities and
cytokine responses in different dose-dependent manner in the immunized mice
model, respectively. These results provided important orientations for the
comprehensively understanding and study of the humoral central immune system
of human, and provided a novel insight on the treatment of serious disease and
immune improvement of human.

5. Peptides. 2011. Epub 2011 April 28.

Isolation, antiproliferation on tumor cell and immunomodulatory
activity of BSP-I, a novel bursal peptide from chicken humoral
immune system.

Author information: Feng X, Liu T, Wang F, Cao R, Zhou B, Zhang Y, Mao X, Chen
P, Zhang H.

Division of Key Lab of Animal Disease Diagnosis and Immunology of China's

Department of Agriculture, Nanjing Agriculture University, Nanjing 210095, China.
Abstract
The bursa of Fabricius (BF) is acknowledged as central humoral immune organ
unique to birds. Our purpose was to identify the potential function of a novel bursal-
derived bioactive peptide. A bursal septpeptide (BSP-I), EPASGMM, first isolated
from BF, reduced MCF and Hela tumor cells proliferation, and enhanced antitumor
factor p53 luciferase activity and protein expression. Further, we found the
significantly immune inducing function of BSP-I on antigen-specific immune
response in BALB/c mice intraperitoneally immunized with inactivated avian
influence virus (AIV, H(9)N(2) subtype) vaccine, including of enhancing the
antibody (IgG, the isotypes IgG1 and IgG2a) production, and stimulating cytokines
IL-4 and IFN-γ level, and inducing T cell immunophenotyping and lymphocyte
proliferation. These results suggested that as the bioactive peptide from avian
humoral immune system, various biological function of BSP-I may have far-
reaching implication on immune system significance, which might provide novel
insight on linking between humoral immune system and development of effective
immunotherapeutic strategies for treating human cancers diseases.

6. Methods Mol Biol. 2014 august 17.

Detection of cell-mediated immune response to avian influenza
viruses.

Author information: Kapczynski DR.

Abstract
The measurement of cell-mediated immunity (CMI) is critical to understanding the
role and regulation of avian lymphocytes following avian influenza virus (AIV)
infection. While these different cell types have distinctly different modes of action in
terms of contributions to protection, they account for the majority of adaptive
immunity induced following infection or vaccination. Although the ability to measure
CMI has steadily improved over the last few years, few studies have examined its
role in protection of birds against AIV. The increasing availability of monoclonal
antibodies recognizing various avian cell-associated antigens has made this
technique more specific and informative.
7. Mol Immunol. 2013 Dec Epub 2013 August 1.
Glycans from avian influenza virus are recognized by chicken
dendritic cells and are targets for the humoral immune response
in chicken.

Author information: De Geus ED, Tefsen B, van Haarlem DA, van Eden W, van

Die I, Vervelde L.

Department Infectious Diseases & Immunology, Faculty of Veterinary Medicine.

Abstract
To increase our understanding of the interaction between avian influenza virus and
its chicken host, we identified receptors for putative avian influenza virus (AIV)
glycan determinants on chicken dendritic cells. Chicken dendritic cells (DCs) were
found to recognize glycan determinants containing terminal αGalNAc, Galα1-3Gal,
GlcNAcβ1-4GlcNAcβ1-4GlcNAcβ (chitotriose) and Galα1-2Gal. Infection of chicken
dendritic cells with either low pathogenic (LP) or highly pathogenic (HP) AIV results
in elevated mRNA expression of homologs of the mouse C-type lectins DEC205
and macrophage mannose receptor (MMR), whereas expression levels of the
human dendritic cell-specific intercellular adhesion molecule-3-grabbing non-
integrin (DC-SIGN) homolog remained unchanged. Following uptake and
subsequent presentation of avian influenza virus by DCs, adaptive immunity,
including humoral immune responses are induced. We have investigated the
antibody responses against virus glycan epitopes after avian influenza virus
infection. Using glycan micro-array analysis we showed that chicken contained
antibodies that predominantly recognize terminal Galα1-3Gal-R, chitotriose and
Fucα1-2Galβ1-4GlcNAc-R (H-type 2). After influenza-infection, glycan array
analysis showed that both levels and repertoire of glycan-recognizing antibodies
decreased. However, analysis of the sera by ELISA indicated that the levels of
different isotypes of anti-glycan Abs against specific glycan antigens was increased
after influenza-infection, suggesting that the presentation of the glycan antigens
and iso-type of the Abs are critical parameters to take into account when
measuring anti-glycan Abs. This novel approach in avian influenza research may
contribute to the development of a broad spectrum vaccine and improves our
mechanistic understanding of innate and adaptive responses to glycans.
8. J Immunol. 2014. Epub 2014 Feb 24.
Ligation of surface Ig by gut-derived antigen positively selects
chicken bursal and peripheral B cells.

Author Information:Davani D, Pancer Z, Ratcliffe MJ.

Abstract
In many mammals and birds, B cell lymphopoiesis takes place in GALT, such as
the avian bursa of Fabricius. Although BCR expression is sufficient for bursal
colonization, the role of BCR ligation in the later stages of bursal B cell
lymphopoiesis remains elusive. To address this directly, we introduced a surface
Ig-related construct with defined Ag specificity containing the Ag-binding portion of
a lamprey variable lymphocyte receptor specific for PE fused to a truncated
chicken μ-chain (VLR(PE)Tμ) into developing chick embryos. VLR(PE)Tμ
expression supports bursal follicle colonization, clonal expansion, and Ig V gene
diversification. VLR(PE)Tμ-expressing B cells migrate to the periphery in the
absence of the Ag starting from day 18 of embryogenesis. VLR(PE)Tμ-expressing
B cells declined rapidly in the bursa and periphery in the absence of Ag after hatch;
however, intrabursal injection of PE prolonged survival of VLR(PE)Tμ(+) bursal and
peripheral B cells. Intrabursal introduction of Ag increased emigration of short-lived
LT2(+) B cells. Peripheral VLR(PE)Tμ(+) B cells were maintained following
intrabursal PE application and contained both short-lived LT2(+) and long-lived
LT2(-) B cells. In the chicken bursa, the later stages of B cell development occur in
the presence of gut-derived Ag; therefore, we conclude that Ag-mediated ligation of
BCR in bursal B cells acts to positively select bursal B cells into both short-lived
and long-lived peripheral B cell populations.

9. Copyright © 2015 Elsevier Ltd. All rights reserved.

Systemic immune responses to an inactivated, whole H9N2 avian
influenza virus vaccine using class B CpG oligonucleotides in
chickens
Author information:
Department of Pathobiology, Ontario Veterinary College, University of Guelph,
Guelph, ON, Canada N1G 2W1.

Abstract

Commercial vaccines against avian influenza viruses (AIV) in chickens consist

mainly of inactivated AIV, requiring parenteral administration and co-delivery of an
adjuvant. Limitations in T helper 1 or T helper 2 biased responses generated by
these vaccines emphasize the need for alternative, more efficacious adjuvants.
The Toll-like receptor (TLR) 21 ligand, CpG oligodeoxynucleotides (ODN), has
been established as immunomodulatory in chickens. Therefore, the objective of
this study was to investigate the adjuvant potential of high (20 μg) and low (2 μg)
doses of CpG ODN 2007 (CpG 2007) and CpG ODN 1826 (CpG 1826) when
administered to chickens with a formalin-inactivated H9N2 AIV. Antibody
responses in sera were evaluated in 90 specific pathogen free (SPF) chickens after
intramuscular administration of vaccine formulations at 7 and 21 days post-hatch.
Antibody responses were assessed based on haemagglutination inhibition (HI) and
virus neutralization (VN) assays; virus-specific IgM and IgY antibody responses
were evaluated by ELISA. The results suggest that the vaccine formulation
containing low dose CpG 2007 was significantly more effective at generating
neutralizing (both HI and VN) responses than formulations with high or low doses
of CpG 1826 or high dose CpG 2007. Neutralizing responses elicited by low dose
CpG 2007 significantly exceeded those generated by a squalene-based
adjuvanted vaccine formulation during peak responses. A significantly higher IgM
response was elicited by the formulation containing low dose CpG 2007 compared
to high and low doses of 1826. Although the low dose of CpG 2007 elicited a
higher IgY response than CpG 1826, the difference was not statistically significant.
In conclusion, 2 μg of CpG 2007 is potentially promising as a vaccine adjuvant
when delivered intramuscularly with inactivated H9N2 virus to chickens. Future
studies may be directed at determining the mucosal antibody responses to the
same vaccine formulations.

10. Accepted 16 September 2013, Available online 25 September 2013

Identification of a linear B-Cell epitope on avian reovirus protein
sigmaC

Author information: Chun-hong Yin, Li-ting Qin, Mei-yu Sun, Yu-long Gao, Xiao-le
Qi, Hong-lei Gao, Yong-qiang Wang, Li-li Jang, Xiao-mei Wang.

Division of Avian Infectious Diseases, State Key Laboratory of Veterinary

Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of
Agricultural Sciences, Harbin 150001, PR China
Abstract

SigmaC (σC) protein, which mediates virus attachment to target cells, is the most
variable proteins of avian reovirus (ARV). It is responsible for inducing protective
antibody immune responses in animals. To understand the antigenic determinants
of σC protein, a set of partially overlapping and consecutive peptides spanning σC
were expressed and then screened with the monoclonal antibody (mAb) 2B5
directed against σC. The mAb 2B5 recognized peptides with the σC
motif 45ELLHRSISDISTTV58. Further identification of the displayed B-cell epitope
was conducted with a set of truncated peptides expressed as GST fusion proteins.
The Western blot and ELISA results indicated that 45ELLHRSISDI54 was the
minimal determinant of the linear B-cell epitope. Using sequences analysis, we
found that this epitope was not a common motif shared among the other members
of the ARV and DRV groups. Furthermore, cross reactivity analysis showed that
the associated coding motif of other ARV and DRV groups was not recognized by
2B5. These data suggested that 45ELLHRSISDI54 was a type-specific linear B-cell
epitope of avian reovirus. The results in this study may have potential applications
in the development of diagnostic techniques and epitope-based marker vaccines
against ARV, which is prevalent in China.

11. Accepted 23 March 2015, Available online 6 April 2015

Molecular detection of the avian malaria parasite Plasmodium

gallinaceum in Thailand 

Author information: Sittiporn Pattaradilokrata, Wisawa Tiyamaneea, Phumin

Simpalipana, Morakot Kaewthamasornb, Tawee Saiwichaic, Jian Lid, Pongchai
Harnyuttanakorna

Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok

Abstract

Avian malaria is one of the most common veterinary problems in Southeast Asia.
The standard molecular method for detection of the avian malaria parasite involves
the phenol–chloroform extraction of parasite genomic (g)DNA followed by the
amplification of parasite gDNA using polymerase chain reaction (PCR). However,
the phenol–chloroform extraction method is time-consuming and requires large
amounts of samples and toxic organic solvents, thereby limiting its applications for
parasite detection in the field. This study aimed to compare the performance of
chelex-100 resin and phenol/chloroform extraction methods for the extraction
of Plasmodium gallinaceumgDNA from whole avian blood that had been dried on
filter papers (a common field sampling method). The specificity and sensitivity of
PCR assays for P. gallinaceumcytochrome B (cytb) and cytochrome oxidase
subunit I (coxI) gene fragments (544 and 588 bp, respectively) were determined,
and found to be more sensitive with gDNA extracted by the chelex-100 resin
method than with the phenol/chloroform method. These PCR assays were also
performed to detect P. gallinaceum in 29 blood samples dried on filter papers from
domestic chickens in a malaria endemic area, where the reliable identification of
seven field isolates of P. gallinaceum was obtained with an accuracy of 100%. The
analysis of cytb and coxI gene nucleotide sequences revealed the existence of at
least two genetically distinct populations of P. gallinaceum in Thailand, both of
which differed from the reference strain 8A of P. gallinaceum. In conclusion, the
chelex-100 resin extraction method is a simple and sensitive method for isolating
gDNA from whole avian blood dried on filter paper. Genomic DNA extracted by the
chelex method could subsequently be applied for the PCR-based detection of P.
gallinaceum and DNA sequencing. Our PCR assays provide a reliable diagnostic
tool for molecular epidemiological studies of P. gallinaceum infections in domestic
chickens and wild birds.

12. Accepted 10 January 2015, Available online 19 January 2015

Identification of morphological differences between avian
influenza A viruses grown in chicken and duck cells

Author information: Firas Al-Mubaraka, b, Janet Dalya, Denise Christiec, Donna

Fountaina, Stephen P. Dunhama.
Department of Microbiology – Virology, College of Veterinary Medicine and
Science, Basra University, Iraq
Abstract

Although wild ducks are considered to be the major reservoirs for most influenza A
virus subtypes, they are typically resistant to the effects of the infection. In contrast,
certain influenza viruses may be highly pathogenic in other avian hosts such as
chickens and turkeys, causing severe illness and death. Following in vitro infection
of chicken and duck embryo fibroblasts (CEF and DEF) with low pathogenic avian
influenza (LPAI) viruses, duck cells die more rapidly and produce fewer infectious
virions than chicken cells. In the current study, the morphology of viruses produced
from CEF and DEF cells infected with low pathogenic avian H2N3 was examined.
Transmission electron microscopy showed that viruses budding from duck cells
were elongated, while chicken cells produced mostly spherical virions; similar
differences were observed in viral supernatants. Sequencing of the influenza
genome of chicken- and duck-derived H2N3 LPAI revealed no differences,
implicating host cell determinants as responsible for differences in virus
morphology. Both DEF and CEF cells produced filamentous virions of equine
H3N8 (where virus morphology is determined by the matrix gene). DEF cells
produced filamentous or short filament virions of equine H3N8 and avian H2N3,
respectively, even after actin disruption with cytochalasin D. These findings
suggest that cellular factors other than actin are responsible for the formation of
filamentous virions in DEF cells. The formation of elongated virions in duck cells
may account for the reduced number of infectious virions produced and could have
implications for virus transmission or maintenance in the reservoir host.

13. Copyright © 2015. Published by Elsevier B.V.

Evaluation of different inactivation methods for high and low
pathogenic avian influenza viruses in egg-fluids for antigen
preparation.

Author information:
National Institute of Virology-Microbial Containment Complex, 130/1, Sus Road,
Pashan, Pune 411021, India
Abstract

In view of the emerging avian influenza (AI) viruses, it is important to study the
susceptibility of AI viruses to inactivating agents for preparation of antigens and
inactivated vaccines. The available information on susceptibility of both the high
and low pathogenic AI viruses to different inactivating agents is inadequate and
ambiguous. It has been shown that different subtypes of influenza viruses require
different physical and chemical conditions for inactivation of infectivity. The present
study was undertaken to evaluate the use of beta-propiolactone (BPL), formalin
and ether for inactivation and its impact on antigenicity of AI viruses.

A total of nine high and low pathogenic AI viruses belonging to four influenza A
subtypes were included in the study. The H5N1 viruses were from the clades 2.2,
2.3.2.1 and 2.3.4. The H9N2 virus included in the study was of the G1 genotype,
while the H11N1 and H4N6 viruses were from the Eurasian lineage. The viruses
were treated with BPL, formalin and with ether. The confirmation of virus
inactivation was performed by two serial passages of inactivated viruses in
embryonated chicken eggs.

The infectivity of all tested AI viruses was eliminated using 0.1% BPL and 0.1%
formalin. Ether eliminated infectivity of all tested low pathogenic AI viruses;
however, ether with 0.2% or 0.5% Tween-20 was required for inactivation of the
highly pathogenic AI H5N1 viruses. Treatment with BPL, ether and formalin
retained virus hemagglutination (HA) titers. Interestingly ether treatment resulted in
significant rise in HA titers (P < 0.05) of all tested AI viruses. This data
demonstrated the utility of BPL, formalin and ether for the inactivation of infectivity
of AI viruses used in the study for the preparation of inactivated virus antigens for
research and diagnosis of AI.

14. Accepted 19 October 2012, Available online 22 November 2012

Association of the chicken MHC B haplotypes with resistance to
avian coronavirus
Author information: Ghida R. Banata , Suzana Tkalcica, , Jennifer A. Dzielawab ,
Mark W. Jackwoodc, , Miguel D. Saggesea, , Linda Yatesd, Renee Kopulosd, W.E.
Brilesd, ,
College of Veterinary Medicine, Western University of Health Sciences, College of
Veterinary Medicine, Texas A&M University.

Abstract

Clinical respiratory illness was compared in five homozygous chicken lines,

originating from homozygous B2, B8, B12 and B19, and heterozygous B2/B12
birds after infection with either of two strains of the infectious bronchitis virus (IBV).
All chickens used in these studies originated from White Leghorn and Ancona
linages. IBV Gray strain infection of MHC homozygous B12 and B19 haplotype
chicks resulted in severe respiratory disease compared to chicks with B2/B2 and
B5/B5 haplotypes. Demonstrating a dominant B2 phenotype, B2/B12 birds were
also more resistant to IBV. Respiratory clinical illness in B8/B8 chicks was severe
early after infection, while illness resolved similar to the B5 and B2 homozygous
birds. Following M41 strain infection, birds with B2/B2 and B8/B8 haplotypes were
again more resistant to clinical illness than B19/B19 birds. Real time RT-PCR
indicated that infection was cleared more efficiently in trachea, lungs and kidneys
of B2/B2 and B8/B8 birds compared with B19/B19 birds. Furthermore, M41
infected B2/B2 and B8/B8 chicks performed better in terms of body weight gain
than B19/B19 chicks. These studies suggest that genetics of B defined haplotypes
might be exploited to produce chicks resistant to respiratory pathogens or with
more effective immune responses.

15. Copyright 2010 Elsevier Inc. All rights reserved.

Non-replicating adenovirus vectors expressing avian influenza

virus hemagglutinin and nucleocapsid proteins induce chicken
specific effector, memory and effector memory CD8 + T
lymphocytes
Author information:

Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M

University, College Station, TX 77843, USA.
Abstract

Avian influenza virus (AIV) specific CD8 + T lymphocyte responses stimulated by
intramuscular administration of an adenovirus (Ad) vector expressing either HA or
NP were evaluated in chickens following ex vivo stimulation by non-professional
antigen presenting cells. The CD8+ T lymphocyte responses were AIV specific,
MHC-I restricted, and cross-reacted with heterologous H7N2 AIV strain. Specific
effector responses, at 10 days post-inoculation (p.i.), were undetectable at 2 weeks
p.i., and memory responses were detected from 3 to 8 weeks p.i. Effector memory
responses, detected 1 week following a booster inoculation, were significantly
greater than the primary responses and, within 7 days, declined to undetectable
levels. Inoculation of an Ad-vector expressing human NP resulted in significantly
greater MHC restricted, activation of CD8 + T cell responses specific for AIV.
Decreases in all responses with time were most dramatic with maximum activation
of T cells as observed following effector and effector memory responses.

16. Copyright © 2012 Elsevier Ltd. All rights reserved.

Horse IgG- and ostrich IgY-F(ab′)2 groups have different affinities

for mice erythrocytes and lymphocytes. Implications for avian
immunoglobulin therapeutic usefulness

Author information:

On Cellular Neuropharmacology Laboratory, Center for Biophysics and

Biochemistry, Venezuelan Institute of Scientific-IVIC

Abstract

We used high sensitivity and resolution fluorescence microscopy to study the

interaction of ostrich IgY, horse F(ab′)2 and horse IgG with mice lymphocyte and
erythrocyte plasma membrane. The immunoglobulins were labeled with fluorescein
isotiocyanate (FITC). Our results show an interaction of IgY with lymphocyte
plasma membrane which does not result in endocytosis of the labeled protein.
Less IgG and its F(ab′)2 fraction bind to lymphocytes, and this binding seems to be
followed by endocytosis producing a diffuse cytoplasmic fluorescence in most
lymphocytes exposed to FITC-IgG or FITC-F(ab′) 2. Cytoplasmic fluorescence
resembling FITC was not observed in lymphocytes exposed to FITC-IgY.
Receptors in the erythrocyte membrane also differentiate between avian and horse
Ig; while erythrocytes exposed to horse Igs became intensely fluorescent for at
least 5 h, no erythrocyte labeling occurred when FITC-IgY was used. Our results
suggest that IgY may be a stronger activator of adaptive immunity than horse IgG
in mammals. Adaptive immunity against IgY is detrimental to its IV therapeutic use
in humans and other mammals.

17. Available online March 2, 2012. PUBLISHED MARCH 19, 2012

Modelling the dynamics of antibody production in BIRDS

Author Information: ANDRES OLARTE, CARLOS CLAVIJO, HERNANDO DÍAZ

Dyna, Year 78, Nro. 166, pp. 174-182. April, 2011. ISSN 0012-7353

Abstract

This paper presents a dynamical model of avian immune response, for antibody
production purposes. Antibodies are fundamental tools for research, diagnosis, and
the treatment of several diseases. It has been recognized for some time that
antibodies obtained from poultry are an effi cient alternative for applications in
medicine and biology. Therefore, a dynamical model for this process might be very
useful. The model being proposed consists of seven non-linear ordinary differential
equations with constant coeffi cients that represent the main cellular and molecular
populations in avian immune response. Values for the parameters were obtained
from avian and mammalian literature. In silico-generated responses in terms of
antibody concentrations are presented and compared to reported kinetics.

18. J Immunol. 2014. Epub 2014 Feb 10.

Negative selection of self-reactive chicken B cells requires B cell
receptor signaling and is independent of the bursal
microenvironment.

Author Information:Davani D, Pancer Z, Cheroutre H, Ratcliffe MJ.

Abstract
Although the negative selection of self-reactive B cells in the bone marrow of
mammals has been clearly demonstrated, it remains unclear in models of gut-
associated B cell lymphopoiesis, such as that of the chicken (Gallus gallus). We
have generated chicken surface IgM-related receptors in which the diversity region
of the lamprey variable lymphocyte receptor (VLR) has been fused to the C region
of chicken surface IgM (Tμ). Expression of a VLR:Tμ receptor with specificity for
PE supported normal development of B cells, whereas a VLR:Tμ receptor specific
to hen egg lysozyme (a self-antigen with respect to chicken B cells) induced, in
vivo, complete deletion of VLR(HEL)Tμ-expressing B cells. In ovo i.v. injection of
PE resulted in deletion of VLR(PE)Tμ-expressing Β cells in the embryo spleen,
demonstrating that negative selection was independent of the bursal
microenvironment. Although chickens transduced with a murine CD8α:chicken Igα
fusion protein contained B cells expressing mCD8α:chIgα, cotransfection of the
mCD8α:chIgα construct, together with thymus leukemia Ag (a natural ligand for
mCD8α), resulted in reduced levels of mCD8α:chIgα-expressing B cells in inverse
proportion to the levels of thymus leukemia Ag-expressing cells. Deletion of
mCD8α:chIgα-expressing cells was specific for B cells and required active
signaling downstream of the mCD8α:chIgα receptor. Ag-mediated negative
selection of developing chicken B cells can therefore occur independently of the
bursal microenvironment and is dependent on signaling downstream of the BCR.

19. Rev. Bras. Cienc. Avic. vol.16 no.2 Campinas April./June 2014

The effect of arginine dietary supplementation in broiler breeder

hens on offspring humoral and cell-mediated immune responses
Author information: Murakami AEI; Silva LMS daI; Fernandes JIMII; Silveira TGVIII;
Garcez Neto AFI

This study was supported by National Council of Technological and Scientific

Development (CNPq), Brazil.

Abstract

The influence of supplementing the diet of broiler breeder hens with arginine (Arg)
on their offspring’s humoral and cell-mediated immune response was evaluated in
two experiments. In experiments I and II, breeder hens were fed diets containing
graded levels of Arg (0.943, 1.093, 1.243, 1.393 and 1.543% digestible Arg). In
experiment I, the offspring was randomly grouped according to the treatment
received by the breeder hens, with five levels of Arg in the maternal diet and six
replicates, giving a total 30 experimental units. In experiment II, the offspring were
grouped in accordance with the treatment received by the breeder hens; however,
Arg was added to the starter diet (1.300, 1.450, 1.600, 1.750 and 1.900%
digestible Arg) and also the growing diet (1.150, 1.300, 1.450, 1.600 and 1.750%
digestible Arg). Supplementation of the broiler breeder hen diet did not influence
(p>0.05) the development of the lymphoid organs (cloacal bursa, thymus and
spleen) of the offspring, whether their diet were supplemented or not.
Nevertheless, greater weight and dimensions cloacal bursa were found in the
supplemented offspring in comparison with the nonsupplemented offspring.
Macrophage phagocytic activity was found to be unaffected (p>0.05),
independently of the Arg supplementation.

The offspring fed with supplemented diets showed a linear reduction in the
antibody titer against Newcastle Disease (p<0.05) at seven days of age, and a
quadratic response (p<0.05) at 28 days of age. The antibody titer in the non-
supplemented offspring was not influenced (p>0.05) by the breeder hen diet. This
study concluded that supplementing the breeder hen diet with arginine is
insufficient to improve the humoral and cellular immune response, requiring
supplementation of the offspring diet.

UDDIN, M. M.; ISLAM, M. S.; BASU, J. & KHAN, M. Z. I. Distribution and

quantification of lymphocytes in the major lymphoid organs of naturally gumboro
infected broilers. Int. J. Morphol., 30(4):1585-1589, 2012.

20. ISSN 1516-635X Oct - Dec 2013 / v.15 / n.4 / 323-328

Distribution and quantification of lymphocytes in the major

lymphoid organs of naturally gumboro infected broilers

Author information: Mohammad Mejbah Uddin; Mohammad Saiful Islam; Joyita

Basu & Md. Zahirul Islam Khan

Abstract

Gumboro disease is caused by the infectious bursal disease virus (IBDV) which
rapidly destroys immature Blymphocytes of bursa of Fabricious, and causes
immune suppression and high mortality in commercial broiler farms in Bangladesh.
To investigate the possible effect of IBDV on lymphocytes and its distribution in the
major lymphoid organs, bursa of Fabricious including spleen and thymus of
naturally Gumboro-infected broilers, a research was conducted in the Department
of Anatomy and Histology, collaboration with the Department of Pathology,
Bangladesh Agricultural University, Bangladesh. Bursa of Fabricious, spleen and
thymus of 21-days-old Gumboro-infected and non-infected broilers of same age
(control) were routinely processed and stained by hematoxylin and eosin to
examine the distribution of lymphocytes in the major lymphatic organs as well as
quantified the number of lymphocytes under high power magnification field and
compared with those of control. The number of lymphocytes in bursa of Fabricious,
spleen and thymus of Gumboro-infected broilers were 27.20 ± 1.53, 66.50 ± 2.70
and 79.30 ± 3.92 whereas 121 ± 3.82, 89.90 ± 2.09 and 106.30 ± 4.07 were in non-
infected control respectively. The numbers of lymphocytes were significantly (p <
0.05) lower in all lymphatic organs of Gumboro-infected broilers than those of non-
infected control. The significant numbers of lymphocytes decrease in spleen and
thymus suggest that IBVD not only destroy lymphocytes in bursa of Fabricious, but
also in spleen and thymus and thus may severely suppress the immune response
of IBVD affected broilers.

21. ISSN 1516-635X Jul - Sept 2013 / v.15 / n.3 / 169-286

Effect of bee pollen on the immunity and tibia characteristics in

broilers

Author Information : Oliveira MC deI, Silva DM daI, Loch FCI, Martins PCII, Dias
DMBI, Simon GA

Abstract

This study was carried out to evaluate the effect of bee pollen (BP) levels on the
IgG and IgM titers, weight of lymphoid organs, and on the tibia morphometric
measures and mineralization in broilers at 21 and 42 days of age. Four hundred
birds were used in an entirely randomized design with four treatments (0, 0.5, 1
and 1.5% of BP feed inclusion) and five replicates. At 21 and 42 days of rearing,
blood samples were collected for IgG and IgM analysis, as well as lymphoid organs
(bursa, thymus and spleen) and the tibiae. There was no effect (p>0.05) of the BP
inclusion on IgG titers, bursa and spleen weights, tibia morphometric measures
and mineralization at 21 and 42 days, IgM titer at 42 days or thymus weight at 21
days. However, IgM titers at 21 days and the thymus weight at 42 days linearly
increased with BP dietary inclusion. It was concluded that up to 1.5% BP can be
included in broiler feeds until 21 days of age to enhance bird immunity .

22. September 12, 2012; Approved: September 9, 2013.

Effect of bacillus subtilis natto on growth performance in

muscovy ducks
Author Information : Sheng-Qiu T a1, Xiao-Ying D ab1, Chun-Mei J a, Jing-Jing P
b, Shan-Shan L b, Jin-Ding C

Abstract

The aim of the present study was to determine whether dietary Bacillus subtilis
natto could affect growth performance of Muscovy ducks. A total of 120 hundred
Muscovy ducks at the age of 1 day were randomly assigned to four groups (30
Muscovy ducks/group), and fed with diets supplemented with 0% (control group),
0.1%, 0.2%, and 0.4% Bacillus subtilis natto, respectively during the 6-week
feeding period. Weight gain, feed intake and feed conversion efficiency of Muscovy
ducks were significantly improved by the dietary addition of Bacillus subtilis natto,
and the results were more significant in 0.4% dietary Bacillus subtilis natto
treatment group; Also, Bacillus subtilis natto reduced Escherichia coli and
Salmonella colonies, and increased lactobacilli population in the ileum and the
cecum. Biochemical parameters, including total protein, GOT (glutamic oxaloacetic
transaminase), GPT (glutamic pyruvic transaminase), AKP (alkaline phosphatase),
triiodothyronine (T3) and tetraiodothyronine (T4) contents (p<0.05) in the serum of
Muscovy ducks were significantly improved (p<0.01), while urea nitrogen, glucose,
triglyceride, total cholesterol concentrations decreased when Bacillus subtilis natto
was added to the diets (p<0.05), and improved duodenum and immune functions.
However, the results above were not significantly different between birds fed 0.1%
Bacillus subtilis natto supplemented diets and the control group (p>0.05). The
results of the present study indicate that diets with 0.4% Bacillus subtilis natto
improved the growth performance of Muscovy ducks by increasing the absorption
of protein, simulating hormone secretion, suppressing harmful microflora, and
improving the duodenal structure and immune functions of Muscovy ducks. It is
suggested that Bacillus subtilis natto is a potential candidate to be used use as a
probiotic to improve the growth performance of Muscovy ducks.
23. Development. 2010. Epub 2010 Aug 11.
Experimental evidence for the ectodermal origin of the epithelial
anlage of the chicken bursa of Fabricius.

Autor Information: Nagy N, Oláh I.

Abstract
The bursa of Fabricius (BF) is a central lymphoid organ of birds responsible for B-
cell maturation within bursal follicles of epithelial origin. Despite the fundamental
importance of the BF to the birth of B lymphocytes in the immune system, the
embryological origin of the epithelial component of the BF remains unknown. The
BF arises in the tail bud, caudal to the cloaca and in close association with the
cloacal membrane, where the anal invagination (anal sinus) of ectoderm and the
caudal endodermal wall of the cloaca are juxtaposed. Serial semi-thin sections of
the tail bud show that the anal sinus gradually transforms into the bursal duct and
proctodeum, which joins the distal part of the cloaca during late embryogenesis.
These anatomical findings raise the possibility that the ectoderm may contribute to
the epithelial anlage of the BF. The expression of sonic hedgehog and its receptor
in the embryonic gut, but not in the BF, further supports an ectodermal origin for
the bursal rudiment. Using chick-quail chimeras, quail tail bud ectoderm was
homotopically transplanted into ectoderm-ablated chick, resulting in quail-derived
bursal follicle formation. Chimeric bursal anlagen were generated in vitro by
recombining chick bursal mesenchyme with quail ectoderm or endoderm and
grafting the recombination into the chick coelomic cavity. After hematopoietic cell
colonization, bursal follicles formed only in grafts containing BF mesenchyme and
tail bud ectoderm. These results strongly support the central role of the ectoderm in
the development of the bursal epithelium and hence in the maturation of B
lymphocytes.

.24. Braz. J. Microbiol. vol.45 no.4 São Paulo Oct./Dec. 2014

Immune response elicited by the oral administration of an

intermediate strain of ibdv in chickens
Author information: Juan Manuel Carballeda, Silvina Chimeno Zoth, Evangelina
Gómez, María Soledad Lucero, María José Gravisaco1 and Analía Berinstein

Biotechnology Institute, National Agricultural Technology Institute, Buenos Aires,

Argentina.
Consejo National Scientific and Technical Research, Buenos Aires, Argentina

Abstract

The immune response elicited by the oral inoculation of an intermediate strain of

infectious bursal disease virus was studied in chickens. A strong over expression
of IL-6, IL-8, IFN and IFN was observed in bursa at 3 days post inoculation
together with an increase in splenic NO2 release. An influx of T-lymphocytes was
also detected.

Key words: Infectious Bursal Disease virus, innate immunity, avian cytokines, flow
cytometry, RT-qPCR.

25. Immunol. 2014 Epub 2014 Feb 24.

Ligation of surface Ig by gut-derived antigen positively selects
chicken bursal and peripheral B cells.

Author Information: Davani D, Pancer Z, Ratcliffe MJ.

Abstract
In many mammals and birds, B cell lymphopoiesis takes place in GALT, such as
the avian bursa of Fabricius. Although BCR expression is sufficient for bursal
colonization, the role of BCR ligation in the later stages of bursal B cell
lymphopoiesis remains elusive. To address this directly, we introduced a surface
Ig-related construct with defined Ag specificity containing the Ag-binding portion of
a lamprey variable lymphocyte receptor specific for PE fused to a truncated
chicken μ-chain (VLR(PE)Tμ) into developing chick embryos. VLR(PE)Tμ
expression supports bursal follicle colonization, clonal expansion, and Ig V gene
diversification. VLR(PE)Tμ-expressing B cells migrate to the periphery in the
absence of the Ag starting from day 18 of embryogenesis. VLR(PE)Tμ-expressing
B cells declined rapidly in the bursa and periphery in the absence of Ag after hatch;
however, intrabursal injection of PE prolonged survival of VLR(PE)Tμ(+) bursal and
peripheral B cells. Intrabursal introduction of Ag increased emigration of short-lived
LT2(+) B cells. Peripheral VLR(PE)Tμ(+) B cells were maintained following
intrabursal PE application and contained both short-lived LT2(+) and long-lived
LT2(-) B cells. In the chicken bursa, the later stages of B cell development occur in
the presence of gut-derived Ag; therefore, we conclude that Ag-mediated ligation of
BCR in bursal B cells acts to positively select bursal B cells into both short-lived
and long-lived peripheral B cell population.