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Studies of experimental infection and cross-immunity between stocks of Cowdria ruminantium in Sokoto red goats.

The course of experimental infection and cross-immunity tests between stocks of Cowdria ruminantium were investigated using eight infected and four uninfected control Sokoto red goats. All eight infected goats reacted to the primary infection. The incubation period varied between 8 and 23 days. During the febrile reaction, the highest rectal temperature recorded was 40 degrees C and the lowest was 39.7 degrees C, compared with a range of 38.6 +/- 0.15 to 38.1 +/- 0.19 in control goats. Following the primary infection, each of the animals was treated with long-acting Tetracycline (Terramycin, Pfizer) at a dose of 20 mg kg-1 body weight to control the resulting temperature reaction. They were reinfected with the homologous stock 3 weeks after treatment to demonstrate whether they had become immune. The homologous challenge did not give any temperature reaction during 3 weeks of monitoring. They were then challenged with the heterologous C. ruminantium stock. This did not result in a febrile reaction during 3 weeks of monitoring. The observed cross-protection indicates antigenic similarity between the two stocks.

Animals↗

Differential dengue cross-reactive and neutralizing antibody responses in BALB/c and Swiss albino mice induced by immunization with flaviviral vaccines and by infection with homotypic dengue-2 virus strains.

We investigated whether cross-reactive and/or cross-protective antibodies against dengue virus could be generated in 6-week-old BALB/c mice by immunization with currently approved flaviviral vaccines, i.e., Japanese encephalitis (JE) BIKEN and yellow fever (YF) 17D. Cross-reactivity with dengue antigens was apparent in at least one-third each of JE-vaccinated mouse sera and of JE/YF-vaccinated mouse sera by dengue enzyme immunoassay, but was not detected in sera of mice immunized with YF vaccine alone. All the immunized BALB/c mice failed to generate neutralizing antibodies against the New Guinea C laboratory (NGC-lab) strain of dengue virus type 2. In addition, we determined the specificity of neutralizing antibodies elicited in 3-week-old Swiss albino mice against two homotypic dengue-2 strains, i.e., NGC-lab and Singapore 1999 (SING/99). Although sera from virus-inoculated mice displayed better neutralization against the corresponding strain, antibodies elicited by NGC-lab exhibited a significantly poorer neutralizing response against the SING/99 strain compared to antibodies elicited by SING/99 against NGC-lab. The differences may be related to sequence variations of approximately 3% between the envelope proteins of both strains. Amino acid disparities at positions 71 (Glu --> Ala), 112 (Ser --> Gly) and 124 (Ile --> Asn), which are found in dengue-2 neutralization escape mutants, were also found in the SING/99 strain. The envelope sequence differences may explain diminished binding of NGC-lab-induced neutralizing antibodies to neutralizing epitopes within the envelope of the SING/99 strain, resulting in a lower titer of neutralizing antibodies against another strain of the same serotype.

Albinism↗

S1 gene sequence analysis of a nephropathogenic strain of avian infectious bronchitis virus in Egypt.

BACKGROUND: Infectious bronchitis is highly contagious and constitutes one of the most common and difficult poultry diseases to control. IBV is endemic in probably all countries that raise chickens. It exists as dozens of serotypes/genotypes. Only a few amino acid differences in the S1 protein of vaccine and challenge strains of IBV may result in poor protection. Tropism of IBV includes the respiratory tract tissues, proventriculus and caecal tonsils of the alimentary tract, the oviduct and the kidney. RESULTS: Infectious bronchitis virus (IBV) strain closely related to Massachusetts (Mass) serotype was isolated from broiler chickens suffering from severe renal and respiratory distresses. The isolate was serologically identified by Dot-ELISA and further characterized by RT-PCR then genotyped using S1 gene sequence analysis. Alignment of the S1 sequence of the isolate with 16 IBV strains revealed high homology to isolates related to Mass serotype. Inoculation with the strain reproduced the disease in experimental 1-day-old chickens and resulted in 20% mortality, severe renal and moderate respiratory distresses. Marked histopathological changes in both kidney and trachea were observed in experimentally infected chickens. A protection study using the H120 live attenuated vaccine showed low protection rate in spite of high S1 sequence homology (97%). Protection based criteria were: virus re-isolation attempts from trachea, tracheal and renal histopathology as well as IBV antigens detection by immunofluorescent antibody technique in kidney sections. CONCLUSION: Periodical evaluation of cross-protective capabilities of IBV vaccine(s) versus recently recovered field isolates should be performed to ensure optimum control of IBV.

Animals↗

Serological comparison and antigenic relationships of seven serotypes of infectious bronchitis virus using the hemagglutination-inhibition test.

The antigenic relationships, antigenic spectrum, and immunogenicity of seven isolates of infectious bronchitis virus (IBV) were examined using the hemagglutination-inhibition (HI) test. Because there was a discontinuity of antigenic relationships and a high degree of cross-reactivity among serotypes of IBV in cross-hemagglutination-inhibition tests, the range of antigenic spectrum used to group the serotypes with the HI test should be wider than the limits suggested by the plaque-reduction test. The HI test may provide valuable information in monitoring the immune status of a flock following vaccination when the area has a history of infectious bronchitis infection. It may also be used as a rapid diagnostic test if a flock is experiencing an outbreak of a disease caused by emergence of a new type of IBV. Interpretation of HI titers in evaluating immune status of chickens following infection with IBV depends on further cross-challenge and cross-protection studies of various types of IBV.

Animals↗

Induction of heterosubtypic immunity to influenza A virus using a DNA vaccine expressing hemagglutinin-C3d fusion proteins.

Cross-protection between different subtypes of influenza A virus has been attributed to heterosubtypic immunity (HSI). Although, HSI can occur in the absence of anti-HA or anti-NA antibodies, HSI seems to be mediated, in part, by cross-reactive antibodies. In this study, we examined the effects of a DNA vaccine expressing an influenza HA fused to three copies of murine C3d of complement (HA-mC3d(3)). HA-mC3d(3) elicited heterosubtypic immunity more efficiently than non-fused forms of HA and protected mice from lethal challenge of influenza with different subtypes. Plasmid encoding for various forms of HA were constructed from two influenza strains, A/Puerto Rico/8/34 (H1N1) or A/Aichi/2/68-x31 (H3N2). Vaccinated mice were analyzed for enhancement of anti-HA titers, affinity maturation of antibody, hemagglutinin-inhibition activity, and altered cytokine profiles. The HA-mC3d(3)-DNA vaccinated mice were protected from heterologous influenza challenge, even though sera from these mice had no viral-neutralizing activity against heterologous virus.

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Characteristics of passive immunity against hantavirus infection in rats.

The protective effects of passively administered antibodies against hantavirus infection were studied in newborn rats. Death as well as infection were completely prevented from intraperitoneal challenge of strain SR-11 (SR) (2 x 10(3) FFU, 10(2.1) LD50), in newborn rats which received 0.1 ml of anti-SR rat serum (neutralizing antibody titer, 1:640) 4 hr before the virus challenge. In these rats, no virus was detected in the peritoneal macrophages, lung, kidney, and brain. The immune serum infusion before the virus challenge also conferred protection to rats against an intramuscular or subcutaneous challenge of strain SR, but did not protect the rats against intracerebral challenge of the virus. In the rats which received the immune serum after the challenge, infection was not prevented, although some of the animals were protected from the death. Virus titers in the lung, kidney, and brain of the rats were reduced by the transfer of the immune serum even as late as 72 hr after the challenge. Cross-protection in the rats which received the immune serum was strong between strains SR and KI-262 within the same serotype, but very weak between strains SR and Hantaan 76-118 of different serotypes.

Animals↗

Dendritic cells pulsed with live and dead Legionella pneumophila elicit distinct immune responses.

Legionella pneumophila is the causative pathogen of Legionnaires' disease, which is characterized by severe pneumonia. In regard to the pathophysiology of Legionella infection, the role of inflammatory phagocytes such as macrophages has been well documented, but the involvement of dendritic cells (DCs) has not been clarified. In this study, we have investigated the immune responses that DCs generate in vitro and in vivo after contact with L. pneumophila. Heat- and formalin-killed L. pneumophila, but not live L. pneumophila, induced immature DCs to undergo similar phenotypic maturation, but the secreted proinflammatory cytokines showed different patterns. The mechanisms of the DC maturation by heat- or formalin-killed L. pneumophila depended, at least in part, on Toll-like receptor 4 signaling or on Legionella LPS, respectively. After transfer to naive mice, DCs pulsed with dead Legionella produced serum Ig isotype responses specific for Legionella, leading to protective immunity against an otherwise lethal respiratory challenge with L. pneumophila. The in vivo immune responses required the Ag presentation of DCs, especially that on MHC class II molecules, and the immunity yielded cross-protection between clinical and environmental strains of L. pneumophila. Although the DC maturation was impaired by live Legionella, macrophages were activated by live as well as dead L. pneumophila, as evidenced by the up-regulation of MHC class II. Finally, DCs, but not macrophages, exhibited a proliferative response to live L. pneumophila that was consistent with their cell cycle progression. These findings provide a better understanding of the role of DCs in adaptive immunity to Legionella infection.

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Antibodies and T cells against synthetic peptides of the C-terminal domain (Hc) of botulinum neurotoxin type A and their cross-reaction with Hc.

Seventeen peptides containing T cell and/or antibody (Ab) epitopes previously localized on Hc of botulinum neurotoxin type A were used in SJL and BALB/c mice as immunogens either individually or as an equimolar mixture of groups that contained epitopes of T cells, Abs or both, to determine their abilities to generate T cells and/or Abs that recognize intact Hc. In SJL, peptide 897-915 which included both T cell and Ab epitopes, elicited Abs that cross-reacted very strongly with Hc. In BALB/c, peptides 869-887, 883-901, 981-999 and 1275-1296 which contained Ab epitopes generated Abs that cross-reacted strongly with Hc. A mixture of peptides that contained T cell and Ab epitopes was effective in both strains in eliciting T cells and Abs that cross-reacted with Hc. This mixture form gave a quicker rise (after two injections) in cross-reactive (with Hc) Ab titer as compared to other peptide mixtures or the individual peptides, and sustained in BALB/c a high Ab titer upon further booster injections. Some of the regions that elicited crossreactive immunity to Hc have sequence similarity to other clostridial toxins, suggesting that one or more of these synthetic peptides might provide cross-protection against those toxins.

Amino Acid Sequence↗

Immunization of crossbred cattle (Bos indicus x Bos taurus) with fractionated midgut antigens against Hyalomma anatolicum anatolicum.

Crossbred calves (Bos indicus x Bos taurus) were immunized with a fractionated midgut supernate antigen (GS-F Ag from Hyalomma anatolicum anatolicum). The first inoculation on day 0 was given intramuscularly after emulsification with Freund's complete adjuvant; the second was given subcutaneously on day 14 in incomplete Freund's adjuvant; and the third on day 35 was given subcutaneously without adjuvant. Each injection comprised 1 mg of antigen protein. Ten days after the last inoculation, the immunized calves were challenged simultaneously with 1000 larvae and 20 pairs (20 males and 20 females) of adult H. a. anatolicum on one ear and a similar number of Hyalomma dromedarii ticks on the other ear. There was a significant decrease in the percentage larval engorgement and larval rejection of up to 34% on the immunized calves. A significant increase in the engorgement and preoviposition periods and a significant decrease in the engorged weight, egg mass weight and reproductive index were observed for adult female ticks when fed on the immunized calves. The GS-F Ag also induced a considerable degree of cross-protection in calves against H. dromedarii larval ticks.

Animals↗

In vitro and in vivo characterization of avian reoviruses. I. Pathogenicity and antigenic relatedness of several avian reovirus isolates.

Pathogenicity, pathogenesis, and antigenic relatedness of four avian reovirus isolates obtained from commercially reared broilers were investigated. Chickens of various ages were inoculated both orally and intratracheally with reovirus. Based on disease signs, mortality, weight depression, tissue lesions, invasiveness, and viral persistence in chickens inoculated at 1 day of age, the isolates were classified as being of low, intermediate, or high pathogenicity. The low-pathogenicity isolate (2177) did not cause mortality, weight depression, or clinical disease. The isolate of intermediate pathogenicity (2035) produced low mortality rates (8%), some weight reduction by 7 weeks postinoculation, and microscopic lesions in the intestine and gastrocnemius tendons. The pathogenic isolates, 2408 and 1733, caused severe clinical disease characterized by stunting, feathering abnormalities, mortality as high as 84%, and microscopic lesions in the liver, intestine, pancreas, and/or gastrocnemius tendon. Highly pathogenic isolates also persisted longer in tissues of infected birds and elicited a more prompt and prolonged antibody response. Birds inoculated at 1 day or 1 week of age were more susceptible to reovirus-induced disease than birds inoculated at 2 weeks, suggesting an age-associated resistance. All isolates produced mortality with equal frequency in embryos. The isolates characterized were found to be antigenically similar based on cross-neutralization and cross-protection studies.

Age Factors↗

Protection in lambs immunised with Haemonchus contortus gut membrane proteins.

Substantial protection against Haemonchus contortus was induced in lambs by hyperimmunising them with an integral membrane extract of intestines dissected from adult worms. The procedure was effective in young growing animals but did not afford any cross protection against either Ostertagia circumcincta or Nematodirus battus. A serum transfer experiment suggested that the effector mechanism was serum antibody which bound to the brush border membrane of the parasites intestinal cells. Sheep which had acquired immunity to H contortus by previous exposure to a trickle infection did not recognise the gut membrane proteins, which suggested that these are normally hidden from the host.

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Specificity of antigens on UV radiation-induced antigenic tumor cell variants measured in vitro and in vivo.

In vitro exposure of nonimmunogenic murine tumor cells to UV radiation (UVR) generates highly antigenic variants that are immunologically rejected by normal, syngeneic mice. The purpose of this study was to determine whether these antigenic variants cross-react immunologically with the parental tumor and whether the UVR-associated antigen unique to UVR-induced tumors is also present on the variants. Antigenic (regressor) variants and nonimmunogenic (progressor) clones derived from UV-irradiated cultures of the C3H K1735 melanoma and SF19 spontaneous fibrosarcoma cell lines were used to address these questions. In an in vivo immunization and challenge assay, the antigenic variants did not induce cross-protection among themselves, but each induced immunity against the immunizing variant, the parent tumor cells, and nonimmunogenic clones derived from UV-irradiated parent cultures. Therefore, the variants can be used to induce in mice a protective immunity that prevents the growth of the parent tumor and nonimmunogenic clones, but not other antigenic variants. In contrast, immunization with cells of the parental tumor or the nonimmunogenic clones induced no protective immunity against challenge with any of the cell lines. Utilizing the K1735 melanoma-derived cell lines in vitro, T-helper (Th) cells isolated from tumor-immunized mice were tested for cross-reactivity by their ability to collaborate with trinitrophenyl-primed B-cells in the presence of trinitrophenyl-conjugated tumor cells. Also, the cross-reactivity of cytotoxic T-lymphocytes from tumor-immunized mice was assessed by a 4-h 51Cr-release assay. Antigenic variants induced cytotoxic T-lymphocytes and Th activity that was higher than that induced by the parent tumor and nonimmunogenic clones from the UVR-exposed parent tumor and cross-reacted with the parental tumor cells and nonimmunogenic clones, but not with other antigenic variants. Furthermore, upon transplantation, the UVR-induced antigenic variants grew in UV-irradiated and immunosuppressed mice, but not in untreated mice indicating that the variants expressed the determinant recognized by suppressor T-cells present in UV-irradiated mice. These results demonstrate that highly antigenic cells generated by the in vitro exposure of two different murine tumors to UV radiation express a determinant shared with the parental tumor cells and nonimmunogenic clones, a unique variant-specific determinant and the suppressor cell-defined determinant present on UVR-induced tumors. Based on these results, two models are proposed to explain the make-up of the antigenic determinants present on the UVR-induced antigenic variants.

Animals↗

Demonstration of cross-reacting tumor rejection antigens in chemically induced respiratory tract carcinomas in rats.

Experiments were performed to determine whether chemically induced rat respiratory tract carcinomas, which possess considerable immunogenicity, contain cross-reacting antigens. In vivo studies showed that effective cross-protection can be induced with four of the five respiratory tract cancers studied, suggesting that these tumors have common tumor rejection antigens. In vitro cytotoxicity studies with sera from tumor-immune hosts showed cross-reactivity among three of the carcinomas tested.

9,10-Dimethyl-1,2-benzanthracene↗

MHC class I+ and class I- HPV16-associated tumours expressing the E7 oncoprotein do not cross-react in immunization/challenge experiments.

It has been demonstrated repeatedly that a high proportion of tumours derived from MHC class I+ precursors are MHC class I-. Since a major task in immunotherapy strategies for treatment of malignancies is to develop polyvalent tumour vaccines efficient against a broad spectrum of tumours, we have examined whether MHC class I+ cell-based tumour vaccines can cross-protect against homologous MHC class I- tumour challenge and vice versa. For these purposes, we have used two oncogenic cell lines induced independently by co-transfection of murine H-2b cells with E61E7 HPV16 and activated Ha-ras oncogenes, the tumours TC-1 (MHC class I+, HPV16 E7+) and E7+). Surprisingly, it was found that these two tumours do not cross-react, although both of them contain the crucial HPV16-coded tumour rejection antigen E7. Preimmunization with the MHC class I+ tumour did not protect against a subsequent challenge with the MHC class I- tumour and vice versa; however, immunization with the TC-1 tumour could protect syngeneic mice against the TC-1 tumour challenge and, similarly, immunization with the MK16/1/IIIABC tumour could protect mice against the MK16/1/IIIABC tumour challenge. If this finding can also be confirmed as a more general phenomenon with other MHC class I+ and class 1- tumours, it could have serious implications for design of immunotherapeutic vaccines and protocols.

Animals↗

A comparison of some immunological methods for the differentiation of strains of foot-and-mouth disease virus.

Two FMDV strains which had been previously differentiated by complement-fixation were compared by guinea-pig protection test, kinetic neutralization and micro-neutralization tests. It was found that these tests, which have not been previously applied by the methods described, were all capable of FMDV strain differentiation. Similar differences were found by all methods, which suggests that comparisons made by cross-CF, cross-neutralization or cross-protection involve measurement of the same antigen/antibody interactions.

Animals↗

An experimental vaccine providing heterologous protection for Leishmania species in murine model.

Five groups each of 20 BALB/C mice were immunized by heat killed L. donovani promastigotes. Two groups served as control. Each group of mice was subdivided into 2 equal subgroups and challenged with either L. donovani or L. major. The results revealed a high cross reactivity paralleled by a high cross protection between L. donovani and L. major. Heat killed L. donovani promastigotes super-passed other tested SDS-PAGE fractionated and eluted L. d. fractions identified by immune BALB/c mice in providing protection against both L. donovani and L. major. The percentage of protection (reduction in parasite burden) offered by each vaccine was as follows: 80.21%, 36.11%, 9.29%, 33.44% and 17.48% respectively against challenge infection with L. donovani and 3.012 x 10(4), 1.095 x 10(4), 6.054 x 10(2), 2.409 x 10(4), and 3.34 x 10(2), respectively against challenge infection with L. major.

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[Immunobiologic properties of swine rotavirus].

Immunobiological properties of swine rotaviruses were studied. A field isolate of this virus was markedly virulent for newborn colostrum-less piglets. An attenuated strain of swine rotavirus was obtained by serial passages in continuous cell cultures (MA-104 and SPEV). The virus of the 60th passage was nonvirulent for colostrum-less piglets although produced marked seroconversion. Naso-oral immunization of piglets with the attenuated strain prevented the manifestation of clinical disease upon challenge with the virulent swine rotavirus. Besides, the swine rotavirus of the 60th passage showed cross-protective activity against virulent human rotavirus in modelling human rotavirus infection in colostrum-less piglets.

Animals↗

Salmonella DNA adenine methylase mutants elicit protective immune responses to homologous and heterologous serovars in chickens.

Salmonella DNA adenine methylase (Dam) mutants that lack or overproduce Dam are highly attenuated for virulence in mice and confer protection against murine typhoid fever. To determine whether vaccines based on Dam are efficacious in poultry, a Salmonella Dam(-) vaccine was evaluated in the protection of chicken broilers against oral challenge with homologous and heterologous Salmonella serovars. A Salmonella enterica serovar Typhimurium Dam(-) vaccine strain was attenuated for virulence in day-of-hatch chicks more than 100,000-fold. Vaccination of chicks elicited cross-protective immune responses, as evidenced by reduced colonization (10- to 10,000-fold) of the gastrointestinal tract (ileum, cecum, and feces) and visceral organs (bursa and spleen) after challenge with homologous (Typhimurium F98) and heterologous (Enteritidis 4973 and S. enterica O6,14,24: e,h-monophasic) Salmonella serovars that are implicated in Salmonella infection of poultry. The protection conferred was observed for the organ or the maximum CFU/tissue/bird as a unit of analysis, suggesting that Dam mutant strains may serve as the basis for the development of efficacious poultry vaccines for the containment of Salmonella.

Animals↗