Disease Virus Infection In Chickens Research Articles

Transition from acute to persistent infection: Antigen persistence and cytotoxic T cells response in infectious bursal disease virus infected SPF chickens (P6129)

Abstract Described by Cosgrove about a half century ago, infectious bursal disease (IBD) is as an acute, highly contagious viral infection of young chickens, which last for 7 to 10 days and leads to immunosuppression. Here, we are reporting for the first time that in IBD virus (IBDV) -infected, specific pathogen free (SPF) chickens bursa, the virus antigen, macrophages, CD4+, CD8+ cells, Fas, Fas ligand (FasL), and caspase-3 were consistently detected by immunohistochemistry up to 56 days post inoculation (DPI). Two-week-old SPF chickens were inoculated intraocularly with standard challenge strain (STC) using a 104embryo infectious dose. Histopathology of bursal tissues revealed severe lymphoid follicular depletion at DPI 5-28. A distinctive pattern of bursal follicles in the form of small and large follicles was detected from DPI 35 to 56. At 42 and 56 DPI a characteristic clustered pattern of IBDV antigen, macrophages, CD4+, CD8+ cells, Fas, FasL, and caspase-3 were detected in or around the small bursal follicles as compared to large bursal follicles. This is the first report on the persistence of IBDV antigen causing a latent infection, in bursal tissues of SPF chicken. Detection of persistent IBDV infection is highly significant in the elucidation of virus pathogenesis, immunology and epidemiology and it raises many questions that need to be answered in future studies.

Differential expression of Toll-like receptor genes in lymphoid tissues between Marek's disease virus-infected and noninfected chickens

Toll-like receptors (TLR) are trans-membrane sensors recognizing invading microbes. Toll-like receptors play a central role in initiating immune responses against several pathogens. In this study, we investigated the response of TLR and downstream genes to Marek's disease virus (MDV) infection. Forty 1-d-old chicks were randomly divided into 2 groups, with 20 chicks infected with MDV and 20 chicks mock-infected. Four chickens were euthanized respectively from infected and age-matched noninfected groups at 4, 7, 14, 21, and 28 d postinfection (dpi). Bursas, spleens, and thymuses were removed. The differential expression of TLR genes, including TLR3, TLR5, TLR7, TLR15, and TLR21, and downstream genes of TLR7, including MyD88, TRAF3, TRAF6, IFNA, IFNB, and IL6, in lymphoid tissues of MDV-infected and noninfected chickens was determined by real-time PCR. The results showed that the change of TLR genes was different in 3 lymphoid tissues. Expression of TLR7 and MyD88 was upregulated at 14 dpi and downregulated at 28 dpi in MDV-infected compared with noninfected spleens. The TRAF6 and IFNB were upregulated, and TRAF3, IFNA, and IL6 genes showed increasing trends in MDV-infected compared with noninfected spleens at 14 dpi. The expression of TLR3 and TLR15 genes was downregulated in MDV-infected compared with noninfected spleens at 28 dpi. The results indicated that TLR7 and its downstream genes were a response to MDV infection at 14 dpi. However, the function of TLR was impaired when the infection entered the tumor transformation phase. In bursas, TLR3 and TLR15 genes were upregulated at 7 and 4 dpi, respectively. It indicated that TLR3 and TLR15 might be involved in response to MDV infection in bursa at early phases. However, no differential expression of TLR genes was observed between MDV-infected and noninfected thymuses, which indicated that the thymus had little response to MDV infection mediated by TLR.

Microsatellite instability in Marek’s Disease Virus infected primary chicken embryo fibroblasts

BackgroundMarek’s disease virus (MDV), an oncogenic α-herpes virus, causes a devastating disease in chickens characterized by development of lymphoblastoid tumors in multiple organs. Microsatellite instability (MSI), a symptom of defect in DNA mismatch repair function, is a form of genomic instability frequently detected in many types of tumors. However, the involvement of MSI in MDV-infected cells has not been investigated. In this study, we determined the presence and frequency of MSI in primary chicken embryo fibroblasts infected with or without MDV strain in vitro.Results118 distinct microsatellite markers were analyzed by polymerase chain reaction (PCR) in 21 samples. MSI was found in 91 of 118 markers, and 12 out of 118 demonstrated frequency of MSI at ≥ 40%. 27 of 118 microsatellite loci did not show microsatellite instability.ConclusionsThese findings showed that MSI was a real event occurring in primary chicken embryo fibroblasts infected with MDV in vitro as evidenced by the high frequency of MSI, and may be specifically associated with genome alteration of host cells during MDV infected.

Fas/FasL and perforin–granzyme pathways mediated T cell cytotoxic responses in infectious bursal disease virus infected chickens

Infectious bursal disease (IBD) is a highly contagious disease of chickens which leads to immunosuppression. In our previous study it was demonstrated that, possibly, CD4+ and CD8+ T cells may employ perforin and granzyme-A pathway for the clearance of IBDV-infected bursal cells. In this study, we evaluated the cytotoxic T cell responses involving two independently functioning but complementary mechanisms: Fas–Fas ligand and perforin–granzyme pathways in IBDV-infected chickens. As demonstrated previously, infection of chickens with IBDV was accompanied by influx of CD8+ T cells in the bursa and spleen. There was an upregulation in the gene expression of cytolytic molecules: Fas and Fas ligand (FasL), perforin (PFN) and granzyme-A (Gzm-A) in bursal and in the splenic tissues of IBDV inoculated chickens. Additionally, for the first time, we detected Fas, Fas ligand, Caspase-3 and PFN producing CD8+ T cells in the bursa and spleen of IBDV-infected chickens. The infiltration and activation of CD8+ T cells was substantiated by the detection of Th1 cytokine, IFN-γ. These data suggest that T cells may be involved in the clearance of virus from the target organ bursa and peripheral tissues such as spleen. The findings of these studies provide new insights into the pathogenesis of IBD and provide mechanistic evidence that the cytotoxic T cells may act through both Fas–FasL and perforin–granzyme pathways in mediating the clearance of virus-infected cells.

MiRNA expression signatures induced by Marek's disease virus infection in chickens

MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression at the post-transcriptional level. Emerging evidence suggests that differential miRNA expression is associated with viral infection and cancer. Marek's disease virus infection induces lymphoma in chickens. However, the host defense response against Marek's disease (MD) progression remains poorly understood. Here, we utilized microarrays to screen miRNAs that were sensitive to Marek's disease virus (MDV) infection. QRT-PCR analysis confirmed the microarray data and revealed expression patterns of some miRNAs in tumor samples. Chicken miRNA gga-miR-15b, which was reduced in infected susceptible chickens and splenic tumors, controlled the expression of ATF2 (activating transcription factor 2). ATF2 was significantly increased in the same group. Our results indicated that differential expression of miRNA in resistant and susceptible chickens was caused by MDV infection, which effectively influenced protein expression of ATF2. This latter result might be related to Marek's disease resistance/susceptibility.

Perturbations in the antioxidant metabolism during Newcastle disease virus (NDV) infection in chicken

The aim of the present study was to investigate the effect of vitamin E on pro/anti-oxidant status in the liver, brain and heart of Newcastle disease virus (NDV) infected chickens. Activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST) and the levels of reduced glutathione and malonaldehyde were estimated in selected tissues of uninfected, NDV-infected and NDV + vit. E-treated chickens. A significant increase in MDA levels in brain and liver (p < 0.05) was observed in NDV-infected chickens when compared to controls. The activities of SOD, CAT, GPx, GR, GST and levels of GSH were significantly (p < 0.05) decreased in brain and liver of NDV-infected chickens over controls. On the other hand, a significant decreased MDA levels and enhanced antioxidant enzyme activity levels were observed in NDV + vit. E-treated animals compared to NDV-infected chickens. Histopathological studies revealed that liver of NDV infected chicken shows focal coagulation and infiltration of hepatocytes, whereas neuronal necrosis and degeneration of Purkinje cells were observed in brain and moderate infiltration of inflammatory cells was observed in heart. However such histological alterations were not observed in NDV + vit. E-treated animals. The results of the present study, thus demonstrated that antioxidant defense mechanism is impaired after the induction of NDV, suggesting its critical role in cellular injury in brain and liver. Further, the results also suggest that vitamin E treatment will ameliorate the antioxidant status in the infected animals. The findings could be beneficial to understand the role of oxidative stress in the pathogenesis of NDV and therapeutic interventions of antioxidants.

Systems analysis of immune responses in Marek's disease virus-infected chickens identifies a gene involved in susceptibility and highlights a possible novel pathogenicity mechanism.

Marek's disease virus (MDV) is a highly contagious oncogenic alphaherpesvirus that causes disease that is both a cancer model and a continuing threat to the world's poultry industry. This comprehensive gene expression study analyzes the host response to infection in both resistant and susceptible lines of chickens and inherent expression differences between the two lines following the infection of the host. A novel pathogenicity mechanism, involving the downregulation of genes containing HIC1 transcription factor binding sites as early as 4 days postinfection, was suggested from this analysis. HIC1 drives antitumor mechanisms, suggesting that MDV infection switches off genes involved in antitumor regulation several days before the expression of the MDV oncogene meq. The comparison of the gene expression data to previous QTL data identified several genes as candidates for involvement in resistance to MD. One of these genes, IRG1, was confirmed by single nucleotide polymorphism analysis to be involved in susceptibility. Its precise mechanism remains to be elucidated, although the analysis of gene expression data suggests it has a role in apoptosis. Understanding which genes are involved in susceptibility/resistance to MD and defining the pathological mechanisms of the disease gives us a much greater ability to try to reduce the incidence of this virus, which is costly to the poultry industry in terms of both animal welfare and economics.

Allele-specific expression analysis reveals CD79B has a cis-acting regulatory element that responds to Marek's disease virus infection in chickens

Marek's disease (MD) is a T cell lymphoma disease of domestic chickens induced by the Marek's disease virus (MDV), a highly infectious and naturally oncogenic alphaherpesvirus. Enhancing genetic resistance to MD in poultry is an attractive method to augment MD vaccines, which protect against MD but do not prevent MDV replication and horizontal spread. Previous work integrating QTL scans, transcript profiling, and MDV-chicken protein-protein interaction screens revealed 3 MD resistance genes; however, a major challenge continues to be the identification of the other contributing genes. To aid in this search, we screened for allele-specific expression (ASE) in response to MDV infection, a simple and novel method for identifying polymorphic cis-acting regulatory elements, which may contain strong candidate genes with specific alleles that confer MD genetic resistance. In this initial study, we focused on immunoglobulin β (CD79B) because it plays a critical role in the immune response and, more important, is transcriptionally coupled with growth hormone (GH1), one of the previously identified MD resistance genes. Using a coding SNP in CD79B and pyrosequencing to track the relative expression of each allele, we monitored ASE in uninfected and MDV-infected F(1) progeny from reciprocal intermatings of highly inbred chicken lines 6(3) (MD resistant) and 7(2) (MD susceptible). Upon screening 3 tissues (bursa, thymus, and spleen) at 5 time points (1, 4, 7, 11, and 15 d postinfection), we observed that MDV infection alters the CD79B allelic ratios in bursa and thymus tissues at 4 and 15 d postinfection in both mating directions. Our results suggest that CD79B has a cis-acting regulatory element that responds to MDV infection and probably cooperates with GH1 in conferring genetic resistance to MD. This result helps validates the use of ASE screens to identify specific candidate genes for complex traits such as genetic resistance to MD.

Nutrition and immunity: the effects of the combination of arginine and tryptophan on growth performance, serum parameters and immune response in broiler chickens challenged with infectious bursal disease vaccine

To explore the effects of the combination of tryptophan (Trp) and arginine (Arg) on growth performance, serum parameters and immune response of broiler chickens challenged with intermediate plus strain of infectious bursal disease virus vaccine, an in vivo experiment was conducted. A corn–soybean meal-based diet containing different levels of Arg and Trp was used. Cobb500 male broiler chickens from 0 to 49 days of age were subjected to a diet supplemented with the combination of Trp and Arg. Growth performance parameters and serum parameters were measured at 27 and 49 days of age. To evaluate the immunomodulatory effects of the combination of Trp and Arg on the challenged chickens, we measured the serum levels of interferon-α, interferon-γ and immunoglobulin G at 27, 35, 42, and 49 days of age. The results showed that the three evaluated immune system parameters including interferon-α, interferon-γ and immunoglobulin G were significantly enhanced after treatment. This enhancement resulted in the recovery of infectious bursal disease virus-infected chickens compared with controls as confirmed by histopathological examinations. Moreover, serum parameters such as albumin and total protein increased, whereas the treatment decreased (P<0.05) the feed:gain ratio, aspartate amino-transferase, alkaline phosphatase, lactic dehydrogenase, triglyceride and cholesterol. These findings suggest that the combination of Arg and Trp has a regulatory effect on growth performance. Moreover, it modulates the systemic immune response against infectious bursal disease.

Expression of perforin–granzyme pathway genes in the bursa of infectious bursal disease virus-infected chickens

Infectious bursal disease (IBD) is an economically important immunosuppressive disease of chickens. The IBD virus (IBDV) actively replicates in B cells and causes severe bursal damage. Generally, T cells are refractory to infection with IBDV but are known to promote virus clearance. However, the mechanisms of T cell mediated viral clearance are not well understood. In this study, we evaluated the molecular mechanisms of cytotoxic T cell responses in the pathogenesis of IBD in chickens. Infection of chickens with IBDV was accompanied by the infiltration of CD4 + and CD8 + T cells in the bursa. There was an upregulation in the gene expression of important cytolytic molecules; perforin (PFN), granzyme-A (Gzm-A), DNA repair and apoptotic proteins; high mobility proteins group (HMG) and poly (ADP-ribose) polymerase (PARP) in the bursa of Fabricius (BF) whereas expression of NK (natural killer) lysin was downregulated. Importantly, PFN producing CD4 + and CD8 + T cells were also detected in the bursa of IBDV-infected chickens by immunohistochemistry. The Th1 cytokines, IL-2 and IFN-γ expression was also strongly upregulated, suggesting the activation of T cells. The findings of this study highlight the mechanisms of IBD pathogenesis and the role of cytotoxic T cells in the clearance of virus-infected cells.

Expression profiling of genes associated with regulatory functions of T-cell subsets in Marek's disease virus-infected chickens

The environment of tumours caused by Marek's disease virus (MDV) in chickens has been shown to have an immunoregulatory phenotype. The objective of the present study was to examine the expression of key T-regulatory markers during various stages of MDV pathogenesis. Specific-pathogen free (SPF) as well as major histocompatibility complex-defined chickens were infected with the RB1B and JM-16 strains of MDV, respectively. CD4+ and CD8+ T cells from the spleens of infected as well as age-matched controls were sorted by flow cytometry at 4, 10, and 21 days post infection (d.p.i.). The expression of molecules such as CTLA-4, IL-2aR (CD25), PD-1 and PDL-1 was quantified by real-time, quantitative, reverse-transcription polymerase chain reaction. There was an up-regulation of CTLA-4 in CD4+ T cells at 4 d.p.i. The expression of PD-1 was also up-regulated in the CD4+ T-cell subset of SPF birds at 21 d.p.i. Furthermore, the expression of PD-1 was enhanced in CD4+ and CD8+ T cells of genetically susceptible chickens, linking this molecule to susceptibility to disease. The expression of CD25 was down-regulated in both SPF and genetically defined birds after infection. This may be a mechanism through which the virus exerts its immunosuppressive effects. In conclusion, the results of the present study provide more insight into immunomodulatory processes that occur in the lymphoid tissues of MDV-infected chickens.

Expression of Cytotoxicity-Associated Genes in Marek's Disease Virus—Infected Chickens

Cytotoxic host responses to Marek's disease virus (MDV) have been attributed to both natural killer (NK) cells and cytotoxic T lymphocytes (CTLs). However, the mechanisms of cell lysis initiated by these cytotoxic responses during MDV infection are not well defined. Therefore, the current study was aimed at elucidating the molecular mechanisms of host cytotoxic responses to MDV infection by investigating the expression of genes in the cell lysis pathway involving granzyme A. Genes encoding cytolytic proteins, NK lysin, and granzyme A were upregulated during early stages of infection, whereas the genes encoding major histocompatibility complex (MHC) class I and the DNA repair and apoptosis protein, poly(ADP-ribose) polymerase (PARP), were downregulated. These findings shed more light on the mechanisms of host response to MDV infection in chickens.

Glucocorticoids modulate NF-κB-dependent gene expression by up-regulating FKBP51 expression in Newcastle disease virus-infected chickens

FK506-binding protein 51(FKBP51, coded by FKBP5) is a co-chaperone molecule that interacts with the chaperone HSP90 and the glucocorticoid receptor (GR) in an inactive GR complex. It is a negative regulator of glucocorticoid action and is replaced by the positive regulator, FK506-binding protein 52 (FKBP52, coded by FKBP4) when hormone binds to GR, which renders the GR complex active. In this study, we found that the expression of FKBP51 mRNA in 12 organs of Newcastle disease virus (NDV)-infected chickens was robustly induced. The level of corticosterone in NDV-infected chickens was also elevated, approximately 2- to 6.5-fold in the organs compared to non-infected control chickens. The induction of FKBP51 mRNA expression was reproduced by dexamethasone treatment, indicating a role for glucocorticoids in the systemic induction of FKBP51 mRNA expression. In chicken UMNSAH/DF-1 cells, nuclear factor kappaB (NF-κB) was activated in an FKBP51-dependent manner. Regulation of the three NF-κB-dependent, anti-apoptotic genes, bcl-2, bcl-x and bfl-1/ A1 was investigated in UMNSAH/DF-1 cells. Dexamethasone treatment of UMNSAH/DF-1 cells resulted in up-regulation of bcl-2, and down-regulation of bcl-x and bfl-1/ A1. Expression of FKBP51 also resulted in down-regulation of bfl-1/A1, but had no effect on bcl-2 and bcl-x, suggesting the involvement of glucocorticoid-FKBP51-NF-κB signaling in the regulation of expression of bfl-1/ A1 in UMNSAH/DF-1 cells. We observed organ-specific up- or down-regulation of expression of, bcl-2, bcl-x and bfl-1/A1 in NDV-infected and dexamethasone-treated chickens. Differential regulation of bfl-1/A1, bcl-2 and bcl-x upon NDV-infection and dexamethasone treatment suggests that additional factors are involved in the regulation of these genes. These results suggest that systemic elevation of FKBP51 in NDV-infected chickens activates NF-κB, which cooperates with other factors to regulate the expression of NF-κB-dependent genes.

Major histocompatibility complex class I is downregulated in Marek's disease virus infected chicken embryo fibroblasts and corrected by chicken interferon

The major histocompatibility complex (MHC) is a part of the immune system which presents epitopes of intracellular antigens on the cell surface. MHC molecules have receptor–ligand binding affinities with T lymphocytes, permitting the latter to detect foreign intracellular infectious agents. Some pathogens, such as herpesviruses, have developed strategies of evading the host response by MHC. This pressure on the immune system brought, in turn, improvements in the antigen-presenting pathway, for example through the effect of interferon (IFN), which can upregulate MHC expression. The main objective of this work was on the one hand, to determine the abilities of three strains of Marek's disease virus (MDV), a chicken herpesvirus, in interfering with the expression of MHC class I molecules in chicken embryo fibroblasts. On the other hand, we analyzed the ability of IFN to reinstate this important immune capability to the infected cells. Our results show that only an oncogenic serotype 1 strain of MDV (RB1B) was able to markedly decrease MHC class I expression, and that addition of IFN reversed this MDV effect.

Early stages of infectious bursal disease virus infection in chickens detected by in situ reverse transcriptase-polymerase chain reaction

Two infectious bursal disease virus (IBDV) strains were inoculated both intranasally and by eye drop into 5-week-old specific pathogen free chickens. The bursa, the liver, the kidney, the spleen, the thymus, the caecal tonsil and the thigh muscle were harvested at 4, 8, 16, 28, 40, 56, 72, 96 h post-inoculation (p.i.) for IBDV detection by in situ reverse transcriptase-polymerase chain reaction and, at the same time, the pathological changes in these tissues were investigated. A typical positive signal was detected in the liver, the kidney and the spleen of chickens inoculated with the very virulent IBDV H strain at 4 h p.i., but not in the thymus, the caecal tonsil or the thigh muscle until 8 h p.i. Virus was also found in the liver, the spleen, the kidney, the thymus, the caecal tonsils and the muscle of birds inoculated with the cell-adapted Ts strain at 4 h p.i. A positive signal was observed in the bursa later than in the other tissues. The signals increased markedly at 8 h p.i. A decrease in bursal lymphocytes was observed in haematoxylin and eosin stained sections at 28 h p.i. for the H strain and at 40 h p.i. for the Ts strain.

Development and composition of lymphoid lesions in the spleens of Marek's disease virus-infected chickens: Association with virus spread and the pathogenesis of Marek's disease

Changes in lymphocyte distribution in spleens of Marek's disease virus (MDV) infected White Leghorn chickens of line 72 (MD susceptible) and line 61 (MD resistant) were studied by immunocytochemistry. Lymphocytes expressing the MDV antigen pp38 (predominantly B cells) were detected from 4 to 6 days post-inoculation (d.p.i.) but not at or after 8 d.p.i., and were more numerous in line 72. In line 61, infection resulted in depletion of B lymphocytes and an increase in T lymphocytes from 3 to 6 d.p.i., but no change in distribution of these cells. From 8 d.p.i., the B-dependent tissue began to recover and the T cells decreased in number. In line 72, infection caused a dramatic change in lymphocyte distribution, with formation of 'lymphoid lesions'. Diffuse, irregular patches of B lymphocytes, around the capillaries, became surrounded by large aggregates of TCRαβ1+ CD8+ and CD4+ lymphocytes, bordered by a band of TCRγδ+ lymphocytes. From 8 d.p.i., the B-dependent areas partially recovered, while TCRαβ1+ CD4+ and CD8+ lymphocytes, potentially transformed, became extensively scattered throughout the spleen. We conclude that in line 72, replication and spread of MDV is more efficient and T cell responses in early infection are greater, favouring the tumour stage of the disease.

Cytology of feather pulp lesions from Marek's disease (MD) virus-infected chickens and its application for diagnosis and prediction of MD.

Cytological changes of feather pulp lesions (FPL) sampled chronologically from the same specific-pathogen free chickens inoculated with Marek's disease virus serotype 1 (MDV) were examined, comparing with their histological changes. The birds having Marek's disease (MD) lymphomas or nerve lesions exhibited the characteristic lesion changes on the cytological smears of FPL; the initial non-suppurative inflammatory to the late lymphomatous FPL. The birds having neither the MD visceral lymphomas nor the nerve lesions manifested only non-suppurative inflammatory FPL on the cytological smears throughout the experimental periods. Histological evaluation of FPL sampled from the same birds confirmed as above mentioned cytological results. From these results, the cytological evaluation of FPL proved to be an effective diagnostic and prognostic tool in foreseeing MD incidence.

Appearance of T Cells in the Bursa of Fabricius and Cecal Tonsils during the Acute Phase of Infectious Bursal Disease Virus Infection in Chickens

Groups of 3-wk-old specific-pathogen-free chickens were inoculated intraocularly with two virulent and one vaccine strain of infectious bursal disease virus (IBDV). During the acute phase of infection, the bursa of Fabricius and cecal tonsil were examined by immunohistochemistry for IBDV antigen and T cell. With all three viruses examined, the infection resulted in the appearance of viral antigen in the bursa accompanied by the presence of CD3+ cells. The CD3+ cells were predominantly TCR2+ and appeared at the site where viral antigens were present. The CD3+ cells continued to persist in the bursa after most of the IgM+ cells and IBDV antigen-positive cells had disappeared. Similar appearance of CD3+ cells was noted following viral antigen accumulation in the germinal centers of cecal tonsils in virulent IBDV-infected chickens. These results demonstrated that infection with IBDV resulted in extensive viral replication in the bursa and the cecal tonsils with an associated accumulation of T cells. The role these T cells may play in the pathogenesis of IBDV infection remains to be established.

Endothelial MHC class II antigen expression and endarteritis associated with Marek's disease virus infection in chickens.

Experimental Marek's disease virus (MDV) infection in chickens was used to study the early pathogenesis of virus-induced atherosclerosis. Previous investigations using this model have reported the occurrence of atherosclerotic lesions after approximately 7 months postinfection. In this study, a total of 75 susceptible Cornell P-line chickens were inoculated intraperitoneally with the CU-2 strain of MDV at 3 days of age and subsequently perfused for histologic examination. At 2, 4, 8, 13, and 20 weeks postinoculation, the ascending aorta and the brachiocephalic and coronary arteries were evaluated for early changes. Expression of class II major histocompatibility complex (Ia) antigen by the vascular endothelium was demonstrated by indirect immunodetection as early as 2 weeks after virus inoculation. This change was followed by significant thickening of the intimal layer associated with mononuclear cell infiltration. All the arteries examined from the MDV-infected chickens were affected. Preliminary immunohistochemical staining showed the presence of CD3+ CD4+, and CD8+ cells among the infiltrating cells. The results suggest that an immunopathologic mechanism may be involved in the early pathogenesis of MDV-induced atherosclerosis in chickens.

Changes in lymphoid organs and blood lymphocytes induced by vitamin A deficiency and Newcastle disease virus infection in chickens

The effect of vitamin A deficiency in the presence or absence of Newcastle disease virus infection (NDV, La Sota strain) on weight of lymphoid organs and on the number and type of circulating white blood cells (WBC) was investigated in chickens. Day-old chickens with limited vitamin A reserves were fed purified diets containing either marginal ( ad libitum) or adequate (pair-fed) levels of vitamin A and at 21–28 days of age; half the chickens in each group were infected with NDV. Vitamin A deficiency resulted only in significantly lower absolute and relative weights of bursa of Fabricius and after infection both weights of bursa and thymus were significantly lower. Relative weight of spleen was significantly higher after infection irrespective of vitamin A status. Liver weights were not affected by vitamin A status and/or NDV infection. Both vitamin A deficiency and NDV infection resulted in lymphopenia, while the lowest number of WBC were observed in vitamin A-deficient chickens during the acute phase of NDV (5 days after infection). Subsequent to lymphopenia due to NDV infection, a marked lymphocytosis was observed in controls and to a lesser extent in vitamin A-deficient birds. These results indicate that vitamin A deficiency, which is aggravated by concomitant NDV infection, affects lymphoid cell systems.