Assessment of the immune response to duck plague vaccinations

An experiment was conducted to assess the immune responses of ducks to duck plague (DP) vaccinations employing one commercial and one laboratory-adapted (LA) DP vaccines. Virus neutralisation and leucocyte migration-inhibition tests were conducted at regular intervals before and after vaccinations. Similarly, ducks in vaccinated and control groups were subjected to challenge infection with virulent DP virus.The commercial vaccine yielded a poor immune response and partial protection on challenge whereas satisfactory responses were obtained in ducks receiving two doses of LA vaccine. The humoral as well as cellular factors were stimulated indicating possible involvement of both the immune responses in the protection from duck plague.     

A sequential histologic and immunohistochemical study of duck hepatitis B virus infection in Pekin ducks

Twenty-nine Pekin ducks were inoculated with duck hepatitis B virus (DHBV), DHBV-free serum, or saline at 1 day of age. Congenitally DHBV-infected ducks were also studied. Ducks were killed periodically during a 92-week study and examined histologically and immunohistochemically to assess liver and extrahepatic inflammation and to detect and characterize DHBV core antigen tissue distribution. DHBV infection produced an asymptomatic but persistent DHBV viremia in all ducks associated with a mild to moderate transient hepatic inflammation apparent at 3 to 6 weeks post-inoculation and waning afterwards. DHBV core antigen was detected in hepatocyte cytoplasm at 1 week post-inoculation, and by 3 weeks post-inoculation scattered pancreatic acinar and islet cells also contained viral antigen. Small numbers of mononuclear cells in the splenic white pulp also contained viral antigen. Viral antigen persisted in all of these tissues throughout the duration of the experiment. No inflammation or tissue injury was detected in any of the extrahepatic tissues during the course of DHBV infection. One DHBV-injected duck developed a hepatocellular carcinoma at 88 weeks of age. Isolated patches of neoplastic hepatocytes contained cytoplasmic DHBV core antigen. The results of this study indicate that DHBV, like mammalian hepadnavirus, is capable of producing a persistent infection of the liver and several extrahepatic tissues and suggest that persistent infection may be associated with the development of hepatocellular carcinoma.     

不同感染剂量MDRV对番鸭免疫反应和细胞毒性作用的影响

用不同剂量番鸭呼肠孤病毒(MDRV MW9710株)感染8日龄番鸭后,通过检测血液中淋巴细胞对ConA、LPS的反应和NK细胞、细胞毒T细胞(CTL)的细胞毒性作用,探讨MDRV感染对番鸭免疫细胞功能和细胞毒性作用的影响。结果显示,不同感染剂量MDRV均会抑制番鸭血液淋巴细胞对ConA、LPS的增殖反应,降低NK细胞和CTL细胞的杀伤活性,且影响程度与剂量相关;同时感染番鸭生长缓慢,脾脏肿大,胸腺和法氏囊缩小。上述结果表明,MDRV感染能导致番鸭免疫抑制,且细胞免疫抑制程度与感染病毒量有关。     

Humoral immune response of the duck to duck hepatitis virus: virus-neutralizing vs. virus-precipitating antibodies

Ducks were induced to develop high-level duck hepatitis virus (DHV)-neutralizing antibodies by inculation with a chicken-embryo-adapted DHV via subcutaneous, intramuscular, and intratracheal routes. Administration of the DHV orally in a gelatin capsule failed to stimulate immune response in the ducks. Contact controls of these ducks also remained negative for anti-DHV antibodies. These observations indicated that the DHV administered orally, in gelatin capsule, failed to infect the ducks. None of numerous duck anti-DHV immune sera, with virus-neutralizing activity in the range of 1.8 to 5.57 log10 median- embryo-infective-dose (EID50) neutralization index, developed precipitin lines against a variety of DHV preparations tested in low- and high-ionic-strength agar. The results suggest that the agar-gel immunodiffusion test is unsuitable for serologic testing of duck sera for anti-DHV antibody activity. Virus-neutralizing activity was revealed in both immunoglobulin M (IgM) and IgG classes of sera of actively immunized ducks. Immunodiffusion tests of Sephadex G-200 fractions of 1-day-old duckling sera with monospecific rabbit anti-duck IgM (DIgM) serum failed to detect DIgM. These results demonstrated that the IgM is not being transferred from the dam to the newly hatched ducklings. Seven- and 14-day-old ducks had DIgM in their sera. However, this IgM had no DHV-neutralizing activity, indicating that it was newly developed by the ducklings, which had no active DHV immune response, not having been exposed to DHV.     

番鸭呼肠孤病毒和H9亚型禽流感病毒共感染对番鸭法氏囊和抗体产生的影响

为探讨番鸭呼肠孤病毒(MDRV)和H9亚型禽流感病毒(H9 AIV)共感染对番鸭法氏囊免疫应答能力的影响,本研究将MDRV或/和H9 AIV人工感染8日龄番鸭,观察法氏囊病理组织学变化,检测法氏囊B细胞增殖能力及RE-5 AIV疫苗免疫后抗体变化规律.结果显示:H9 AIV感染组番鸭发病率低(10％),无死亡,法氏囊无病理变化,显著抑制番鸭法氏囊细胞增殖反应;MDRV感染番鸭发病率70％,死亡率40％,生长迟缓,法氏囊病理变化为萎缩,淋巴细胞减少,局部出现范围较小的坏死灶,番鸭法氏囊细胞增殖反应下降;共感染组番鸭发病率100％,死亡率80％,番鸭生长迟缓,法氏囊萎缩和淋巴细胞增殖反应下降程度均比单一病毒感染组严重.病毒感染使番鸭对RE-5 AIV疫苗免疫应答能力明显下降,其共感染组抑制抗体应答程度最严重;共感染组的病毒检出时间早于并且检出率大于单一病毒感染组.表明MDRV与H9 AIV共感染在番鸭免疫应答抑制方面有协同作用.     

MDRV和H9 AIV共感染对番鸭免疫反应的抑制作用

8日龄番鸭人工感染番鸭呼肠孤病毒（MDRV）和/或H9亚型禽流感病毒（H9AIV）,探讨病毒感染对番鸭免疫反应的影响。结果显示,H9AIV感染组番鸭发病率低（10%）,无死亡;显著抑制番鸭淋巴细胞增殖反应,抑制法氏囊和胸腺的发育,脾脏肿大,但不影响生长。MDRV单独感染番鸭发病率75%,病死率45%,对番鸭淋巴细胞增殖...     

Antibody and lymphoblastogenic responses of dogs experimentally infected with Trypanosoma cruzi isolates from North American mammals

The humoral and cellular immune responses of dogs infected with either a non-pathogenic Trypanosoma cruzi isolated from a North American dog (Tc-D) or a pathogenic T. cruzi isolate from an opossum (Tc-O) were studied over a 240 day period. Antibody to T. cruzi epimastigote antigens prepared from Tc-O or Tc-D isolates were first detected by ELISA by Day 26 post infection (PI), peaked by day 175 PI and remained elevated throughout the experimental period in both Tc-O and Tc-D infected dogs. Differences in antibody levels between infected groups were not detected. Western blot analyses were performed using Tc-O and Tc-D epimastigote antigens probed with pooled sera and sera from individual Tc-O and Tc-D infected dogs prior to infection (Day 0), and during the acute (Day 16-35 PI), indeterminate (Day 50-135 PI) and chronic (Day 235 PI) stages of infection. Generally, the patterns, number of protein bands, and temporal appearance of the protein bands identified by pooled sera and sera from individual dogs within each antigen preparation were similar. However, similarities and differences were present in antibody responses between sera from Tc-O and Tc-D infected dogs. Blastogenic responses of peripheral blood mononuclear cells (PBMC) from Tc-O and Tc-D infected dogs to mitogens (concanavalin A, phytohemagglutinin and pokeweed) were not significantly different from controls at any time during the experimental period. The PBMC from both groups of dogs were unresponsive to epimastigote antigens during the acute stage of infection. Statistically significant differences (P less than 0.05) in PBMC responsiveness from controls were observed on Days 70 and 175 PI. Responses decreased to pre-infection levels by Day 240 PI. These studies demonstrate that although two North American T. cruzi isolates have markedly different virulence for dogs, some aspects of their cellular and humoral immune responses are similar while other responses, such as antibody recognition of specific T. cruzi antigens, vary.     

Experimental vesicular stomatitis virus infection of swine: Extent of infection and immunological response

Swine, a natural host species for infection by vesicular stomatitis virus (VSV), were infected with VSV-New Jersey (VSV-NJ) serotype virus obtained from a recent field isolate. Tissues collected from the infected pigs were examined for the presence of infective virus, for viral antigens, and/or for viral nucleic acid. Infective virus could be recovered from tissues near the site of infection for as long as 6 days after the primary infection with VSV. However, no infective virus was recovered following hypothermia induced 11 weeks after infection, or following a secondary challenge with virus 22 weeks after initial infection. Immunofluorescence tests for viral antigens and nucleic acid hybridization assays failed to detect viral antigens or nucleic acids in tissues from which no infective virus could be recovered. Titers of serum-neutralizing antibody peaked 3–5 weeks after infection and then fell slightly until the secondary infection which caused a rapid anamnestic response. Peripheral blood mononuclear cells (PBM) tested 3, 5, 8 or 18 weeks after primary infection all produced readily detectable antigen-specific proliferative responses when cultured with VSV. Thus, although direct tests failed to demonstrate persistence of virus after infection, the humoral and cellular immune response remained elevated for months. Infective VSV was not required to stimulate the proliferative response since UV-inactivated VSV was immunogenic in these in vitro tests. Following primary infection, antigen-specific proliferative responses could be stimulated by several strains of VSV-NJ, but not by VSV-Indiana (VSV-Ind) serotype virus. Secondary infection had relatively little effect on the proliferative response to VSV-NJ strains, but it did cause the PBM to gain responsiveness to VSV-Ind.     

Effect of pasteurella multocida toxin on in vivo immune responses in piglets

Summary

Atrophic rhinitis is characterized by a lack of an immune response towards Pasteurella multocida toxin (Pm‐T). The effects of intranasal administration of Pm‐T on cellular and T‐cell dependent antibody responses of piglets against various other antigens were studied in a 3 by 2 factorial arrangement of treatments: three levels of challenge with Pm‐T (either once; on three consecutive days; or no Pm‐T challenge), and with or without simultaneous immunization with a ‘cocktail’ containing Keyhole limpet haemocyanin (KLH), ovalbumin (OA) and tetanus toxoid (TT). Challenge with Pm‐T affected but did not abolish the in vivo humoral immune response against OA, KLH, or TT. Pigs treated once with Pm‐T showed lower total antibody responses, caused by lower IgG responses to the antigens. We found no clear consistent effects of Pm‐T on cellular immune responses to the various T‐cell‐dependent antigens in vitro. The lack of immune responsiveness to Pm‐T in Pm‐T challenged pigs is discussed. The absence of detectable immune responses does not depend on a general suppression of the cellular immune system.     

Modulation of the cellular immune responses to T-cell-dependent and T cell-independent antigens in lambs with induced bovine viral diarrhea virus infection

Functional interaction between lymphoid cells and lymphotropic viruses is particularly evident for bovine viral diarrhea virus (BVDV) in cattle and its closely related virus, the border disease virus (BVDV) in sheep. The most important aspect of acute or chronic phases of BVDV or BDV infection was the host's increased susceptibility to secondary bacterial or viral infection. To study the ability of BVDV to alter the development of the cellular immune responses to concomitant inoculation with T cell-dependent and T cell-independent antigens, lambs were inoculated twice with rabbit RBC and Escherichia coli lipopolysacharide (LPS) and then were infected with a cytopathic strain of BVDV at postinoculation day 3. Leukopenia characterized by lymphopenia developed after BVDV infection. Increased [3H]thymidine incorporation was observed in resting or lectin-stimulated blood mononuclear cells in the first weeks after inoculation in BVDV-infected lambs, but was followed by decreased [3H]thymidine incorporation after the second inoculation for up to 8 weeks after initial inoculation. In contrast, transient decrease of blastogenic responses, associated with toxic effect of LPS, was detected in inoculated noninfected lambs, but was followed by stimulation of cellular immune responses. Inoculated noninfected lambs had good in vitro cellular immune response to rabbit RBC and LPS antigens, whereas lymphocytes from BVDV-infected lambs could not mount lasting cellular immune responses to antigens or BVDV. Results suggest that BVDV infection in lambs modulates the ability of lymphocytes to respond to lectins or antigenic stimuli according to the time after infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

雏鸭感染鸭肝炎病毒后血液中一氧化氮和一氧化氮合酶的变化

用不同毒力株鸭肝炎病毒人工感染雏鸭，测定雏鸭血浆一氧化氮（NO）和一氧化氮合酶（NOS）的含量，研究NO在鸭病毒性肝炎发病过程中的作用。结果表明：雏鸭感染不同毒力株鸭肝炎病毒后第1天血浆一氧化氮和一氧化氮合酶含量就开始上升，第3天显著或极显著高于对照组，并持续至实验结束。揭示一氧化氮同雏鸭病毒性肝炎的发生发展过程有关。     

Effect of fowl adenovirus-1 (IBH isolate) on humoral and cellular immune competency of broiler chicks

Fowl adenovirus-1 (FAV-1), isolated from field outbreaks of inclusion body hepatitis (IBH), was administered orally to 3-week-old disease-free broiler chicks. Humoral immune competency was evaluated by determining the antibody response of infected chicks to sheep red blood cells (SRBC) and Brucella abortus. FAV-1 infection significantly decreased the antibody response of chicks to B. abortus (T-cell-independent antigen) by decreasing IgM responses, however, the decreased antibody response to SRBC (T-cell-dependent antigen) was statistically non-significant. Bursal index was also found lowered in infected chicks as compared to the control chicks. A significant decrease was seen in blastogenesis response of peripheral blood lymphocytes to phytohaemagglutinin (PHA-P) in FAV-1-infected chicks on 2 and 3 weeks post-infection (WPI). These results indicated that FAV-1 affects humoral as well as cellular immune competency of infected chicks.     

Antibody-mediated resistance against duck enteritis virus infection.

Vaccination of ducks with an apathogenic strain of duck enteritis virus resulted in protection against challenge with the virulent Lake Andes strain of duck enteritis virus by intramuscular inoculation or contact exposure. Antisera produced in the vaccinated ducks were able to transfer resistance against the challenge strain to recipient ducks. Antisera against duck enteritis virus were cytotoxic for duck enteritis virus-infected duck embryo fibroblasts in the presence of guinea pig complement. These in vivo and in vitro data suggest that the humoral immune mechanism plays a role in protecting ducks from duck enteritis virus infection.     

Cell-mediated immune response in hamsters infected with Babesia microti

A study was made of the cell-mediated immune (CMI) response in hamsters infected with Babesia microti isolated from a woman with clinical babesiosis. The leukocyte migration inhibition test (LMIT) using spleen leukocytes exposed to a soluble B. microti antigen was used for in vitro measurement of cellular immunity. The earliest detectable response was noted at the end of the second week after infection and persisted at relatively high levels throughout the 41-week observation period. Challenge infection caused a transitory but strong rise in the LMIT results. No hematologic manifestations of babesiosis were related to the challenge inoculation. The CMI responses were compared wiht humoral antibody measurements using the indirect fluorescent antibody (IFA) test.     

Intestinal mucosal immune response against virulent duck enteritis virus infection in ducklings

Duck virus enteritis (DVE) is an acute and contagious herpes virus infection of duck, geese and swans with high morbidity and mortality. The development of specific mucosal immune system against duck enteritis virus (DEV) infection for ducks has been hindered by a lack of knowledge concerning the purification of immunoglobulin A (IgA) of duck. In the present work, the method for purification of duck immunoglobulin A was developed, and the induction of intestinal mucosal immune responses against DEV was studied by orally infected ducklings with virulent DEV. The results showed that a continuous increased DEV DNA levels were observed in blood and various organs examined of orally infected ducklings throughout the infection, which was accompanied by the development of infection in ducklings from mild progressed to severe pathological lesions. Furthermore, a marked increased level of DEV-specific IgA and IgG antibodies in bile, serum and the intestinal tract, as well as the density of IgA⁺ cells in intestine were detected between 1 and 12 days p.i., followed by a drastic reduction of the antibody levels and the density of IgA⁺ cells at 15 days p.i. The results indicate that the DVE infection can stimulate both IgA-dominated antibody immune responses in the intestinal tract, and IgG-dominated antibody systemic immunity in the serum of ducklings orally inoculated with virulent DEV. The severe lesions of the villus epithelial cells and the lymphoid organs can suppress the intestinal mucosal immune responses.     

Study of immune function in inbred miniature pigs vaccinated and challenged with suid herpesvirus 1.

Specific immune responses of inbred miniature pigs following vaccination and challenge with suid herpesvirus 1 (SHV-1) were determined. Vaccination of swine with SHV-1 elicited both specific neutralizing antibody and lymphoproliferative responses. Moreover, pigs vaccinated with SHV-1 were fully protected against a lethal virus challenge. Pigs vaccinated with a recombinant (r) SHV-1 virus, followed by challenge with a virulent SHV-1, had lower percentages of circulating T- and B-lymphocytes, and showed a significant (P < or = 0.05) reduction in peripheral blood mononuclear cell (PBMC) antibody-dependent cell-cytotoxicity than control (noninfected, SHV-1 sero-negative) animals. From the 5th through the 8th week of postchallenge, rSHV-1 was isolated from 2 of 4 pigs. Presence of r-virus was indicative that PBMC were infectious in vivo. The rSHV-1, with beta-galactosidase activity, was only recovered from ConA- and IL-2-stimulated primary PBMC cocultivated with porcine kidney cells. Control pigs exposed to challenge SHV-1 elicited both specific neutralizing antibody and lympho-proliferative responses followed by subsequent infection. These infected pigs, compared to control pigs, had significantly (P < or = 0.05) lowered percentages of T- and B-lymphocytes, lowered T-cell mitogenic responses, variable PBMC counts, and lowered blood phagocytic cell function. When PBMC from control pigs were cultured and infected with SHV-1, the virus caused a significant (P < or = 0.05) suppression of T-cell proliferation and PBMC mitochondrial dehydrogenase and macrophage activities.     

Muscovy duck reovirus infection rapidly activates host innate immune signaling and induces an effective antiviral immune response involving critical interferons

Muscovy duck reovirus (MDRV) is a highly pathogenic virus in waterfowl and causes significant economic loss in the poultry industry worldwide. Because the host innate immunity plays a key role in defending against virus invasion, more and more attentions have been paid to the immune response triggered by viral infection. Here we found that the genomic RNA of MDRV was able to rapidly induce the production of interferons (IFNs) in host. Mechanistically, MDRV infection induced robust expression of IFNs in host mainly through RIG-I, MDA5 and TLR3-dependent signaling pathways. In addition, we observed that silencing VISA expression in 293T cells could significantly inhibit the secretion of IFNs. Remarkably, the production of IFNs was reduced by inhibiting the activation of NF-κB or knocking down the expression of IRF-7. Furthermore, our study showed that treatment of 293T cells and Muscovy duck embryo fibroblasts with IFNs markedly impaired MDRV replication, suggesting that these IFNs play an important role in antiviral response during the MDRV infection. Importantly, we also detected the induced expression of RIG-I, MDA5, TLR3 and type I IFN in Muscovy ducks infected with MDRV at different time points post infection. The results from in vivo studies were consistent with those in 293T cells infected with MDRV. Taken together, our findings reveal that the host can resist MDRV invasion by activating innate immune response involving RIG-I, MDA5 and TLR3-dependent signaling pathways that govern IFN production.     

Development of a capture ELISA to determine kinetics of soluble CD25 following in vitro and in vivo stimulation of duck peripheral blood monocytes

In humans and other mammals, the α-chain of interleukin-2 (IL-2) receptor (CD25) is induced and expressed on the cell surface after lymphocyte activation and is released from the membrane of activated cells as a smaller soluble form (sCD25). However, little is known about avian sCD25. In the present study, we developed an antigen capture enzyme-linked immunosorbent assay (AC-ELISA) to detect serum sCD25 in ducks, and we used flow cytometry (FCM) to analyze the frequency of CD25(+) cells in the peripheral blood of ducks infected with H9N2 or H5N1 avian influenza virus (AIV) or serotype II Riemerella anatipestifer (RA). Using the AC-ELISA, duck sCD25 molecules were detected in the supernatant and lysates of concanavalin A (Con A)-stimulated peripheral blood mononuclear cells (PBMC), and in the serum of ducks infected with H5N1 virus and RA. However, no sCD25 was detected in the serum of H9N2 AIV-infected ducks. FCM analysis revealed that CD25(+) cells were upregulated within the PBMC of RA-infected ducks throughout the experiment until death, while in the PBMC of H9N2- and H5N1 AIV-infected ducks, the frequency of CD25(+) cells increased in the early stage of infection and then returned to a lower level. Our findings confirm that the dynamics of sCD25 and CD25(+) cells are different in the peripheral blood of ducks infected with H9N2 virus, H5N1 virus, and RA.     

Analysis of immune responses in dogs with canine visceral leishmaniasis before, and after, drug treatment.

The incidence of canine visceral leishmaniasis (CVL) is increasing in the Mediterranean region. Many drugs have been tested for treatment of CVL, but little is known regarding their effect on test immune responses. In our study, three dogs naturally infected with Leishmania infantum and five dogs experimentally infected with the same strain, were treated with dimethasulfonate pentamidine (Lomidine) and the immune response evaluated before, and after, treatment. After the last injection, animals began to gain weight and the major clinical signs disappeared. Antibody titers gradually decreased to low levels, six months after treatment. At the same time, antigen specific lymphoproliferation reappeared in the sampled animals. This study shows that, after treatment, immune cellular responses to leishmanial antigens, involved in protection against Leishmania infection, were established.