Effects of diet, sham-immunization and bleeding on the immune response kinetics for two strains of Cyprinus carpio

The effects of diet and experimental procedures for immunization and bleeding on the immune responsiveness of two strains of Cyprinus carpio were investigated. In addition, the relationship between the genetic structure of these fish, based upon the pattern of scaling, and antibody production was studied. The antibody response to sheep red blood cells (SRBC) was assessed by direct haemagglutination. The control titres for both uninjected and sham-immunized fish, using phosphate buffer saline, were evaluated for each treatment. The results show that diet and multiple bleeding influence the antibody response kinetics to SRBC. The observed differences are dependent upon the strain from which the serum is isolated. Furthermore, the change in antibody titre in the controls is negligible when compared to the increase observed after immunization with SRBC.     

Response and function of cutaneous mucosal and serum antibodies in barramundi (Lates calcarifer) acclimated in seawater and freshwater

Mucosal and serum antibody responses were studied in sibling barramundi (Lates calcarifer) acclimated in either seawater or freshwater following vaccination by intraperitoneal injection or direct immersion in an inactivated Streptococcus iniae vaccine. As expected, route of vaccination had a marked effect on immune response, with direct immersion resulting in low serum antibody levels against S. iniae by ELISA detected 21 days post vaccination at 26 degrees C, whilst a significant response was detected in mucus. A strong specific antibody response was detected in both mucus and serum 21 days following intraperitoneal injection. Fish acclimated in seawater prior to vaccination showed a markedly higher specific mucosal antibody response than sibling fish acclimated in freshwater, regardless of the route of vaccination, whilst the serum antibody response was not affected by salinity. Both mucosal and serum antibodies from fish in seawater and freshwater were capable of binding antigen at salinities similar to full strength seawater in a modified ELISA assay. These results indicate that this euryhaline fish species is not only able to mount significant specific antibody response in cutaneous mucus, but that these antibodies will function in the marine environment.     

Temperature influence on Arctic charr (Salvelinus alpinus L.) antibody response to a cellular antigen

In order to elucidate the immune responses of Arctic charr in relation to temperature, groups were acclimated to a moderate (9°C) and a cold temperature regime (4°C), as well as subjected to a temperature decrease (from 9 to 4°C) immediately prior to an immunization with sheep red blood cells. The charr kept at 9°C responded with increased primary and secondary antibody titres, as seen by direct haemagglutination, while fish at 4°C, as well as the fish subjected to a temperature reduction, displayed lower and lowest antibody titres, respectively, and only after a second immunization. It is concluded that Arctic charr can respond to a cellular antigen with a humoral immune response typical for other teleosts, but that the immune response is delayed and diminished at low temperatures. This temperature-induced immune suppression is intensified if the fish have not been acclimated to cold water prior to immunization. Accepted: 10 October 1999     

Characterization of serum and mucosal antibody responses in white sturgeon (Acipenser transmontanus Richardson) following immunization with WSIV and a protein hapten antigen

Serum and cutaneous mucus antibodies were monitored in white sturgeon for 15 weeks following intraperitoneal immunization. Ten fish were immunized (50 microg) with white sturgeon iridovirus (WSIV) or white sturgeon gonad (WSGO) tissue culture cells emulsified with or without FCA. An additional group was immunized with FITC:KLH+FCA. Fish were booster immunized at 6 weeks. Fish immunized with FITC:KLH+FCA produced significant serum antibodies to FITC by 6 weeks and this response peaked at 12 weeks (average titer 31,000). Mucosal antibodies to FITC were first detected at 12 weeks and significantly elevated by 15 weeks (average titer 18). Anti-WSIV antibody titers were detected in the serum by 9 weeks in fish immunized with WSIV and WSIV+FCA, but only a small number responded to immunization. At 15 weeks, four fish immunized with WSIV produced serum antibodies (average titer 838) and one fish immunized with WSIV+FCA had a serum titer of 1600. Mucosal anti-WSIV antibody titers of 8 and 16 were observed in two fish from the WSIV group at 12 weeks while four different fish from this group responded at 15 weeks (average titer 4). Western Blot using a monoclonal antibody confirmed immunoglobulin in mucus, and specificity to WSIV was further demonstrated by immunocytochemistry using serum from fish immunized with WSIV. Specific antibody was not detected in mucus of fish immunized with WSIV+FCA, WSGO, or WSGO+FCA. Collectively, these experiments demonstrate that white sturgeon can generate a specific antibody response following immunization, and is the first report showing mucosal immunoglobulin is present in this species.     

Variability of immune response in mice immunized at different time intervals after total or partial removal of organs with different reparative capacity]

The antibody response on sheep erythrocytes in the murine spleen at different time intervals after has been studied in one of the following operations: liver resection (70%), unilateral nephrectomy or sialadenectomy, or castration and bilateral sialadenectomy or castration. It was shown, that the liver and kidney surgery enhance the immune response if immunization was performed immediately or 24 h after the operation. Analogous changes of the immune response were observed under the immunization during 3 days after unilateral nephrectomy. The unilateral sialadenectomy evokes the contrary changes in immune response: immunization immediately or 24 h after the operation is accompanied by decrease of the above, the immunization on 3-7 days after the operation gets the increase of antibody genesis. Unilateral and bilateral castration as well as bilateral sialadenectomy evoke no changes of the immune response.     

Cell-mediated immune response to mumps virus infection in man.

The antibody and cell-mediated immune response to mumps virus infection was studied in groups of subjects after natrually acquired mumps virus infection, after parenteral immunization with live attenuated mumps vaccine, and in a population of mumps seronegative subjects. The technique of neutralization of tissue culture infectivity was utilized to study mumps specific antibody. The cell-mediated immunity (CMI) was detected by specific immune release (SIR) of radioactivity by purified lymphocytes after they were reacted with radioactive chromium (51Cr) labeled human conjunctival cell cultures chronically infected with mumps virus. No SIR activity was observed in lymphocytes obtained from cord blood and young individuals seronegative for antibody to mumps virus. Detectable SIR activity was observed in a few older seronegative subjects; however, immunization with mumps vaccine in such antibody negative subjects failed to result in the development of any antibody response in the serum. High SIR activity was observed in the lymphocytes of naturally infected and vaccinated subjects. Although all naturally infected or immunized subjects had varying levels of mumps specific antibody activity in the serum, no correlation existed between the levels of antibody and SIR activity. These observations suggest the development of mumps specific in vitro correlates of CMI after naturally acquired or vaccine-induced mumps virus infection.     

Electron microscopic examination of antigen uptake by salmonid gill cells after bath immunization with a bacterin

Atlantic salmon, Salmo salar , were given 2-min bath immunization with Yersinia ruckeri O-antigen bacterins at doses of 10, 100, and 1000 μg ml⁻¹. The uptake of the antigen was followed by light and electron microscopy of samples taken immediately and periodically after immunization, and the immune response monitored by the passive haemolytic plaque assay. The particulate antigen was observed in the gill mucus, adhering to and within the pavement cells covering the gill filaments, and in mononuclear phagocytes below the epidermal gill cells. There was a direct doseresponse correlation in the observed immune response according to the numbers of splenic antibody-producing cells 14 days after immunization. The cells involved in the recognition and uptake of a bacterin are initial important steps in the immune response, and these studies may aid in the immunopotentiation of fish vaccines and bacterins.     

Role of individual glycoproteins of human parainfluenza virus type 3 in the induction of a protective immune response.

Affinity-purified hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins of human parainfluenza virus type 3 (P13 virus) were used to investigate their role in the induction of a protective immune response following immunization of hamsters. The efficacy of immunization with the glycoprotein antigens was tested by challenge infection. Results of virus recovery from lungs and trachea demonstrated that although immunization with HN or F alone induced an antibody response to the respective glycoproteins, it did not provide a significant level of protection. However, immunization with a mixture of both purified glycoproteins induced higher virus-neutralizing activity in bronchial lavages and afforded complete protection from challenge infection. Similarly, incomplete protection was observed after passive transfer of monospecific rabbit antibody to the purified HN or F in baby hamsters. On the other hand, passive transfer of a mixture of antibodies to HN and F conferred a higher level of protection. Thus, the presence of antibody to both glycoproteins of P13 virus may be essential for protective immunity.     

An analysis of the primary and secondary antibody response in intact and thymectomized rainbow trout, Salmo gairdneri Richardson, to human gamma globulin and Aeromonas salmonicida

The antibody response to human gamma globulin (HGG) and Aeromonas salmonicida (AS) in control and long-term thymectomized rainbow trout was individually monitored in 120 fish. The fish responded poorly to HGG and no memory response was observed. There was no difference in response between control fish and fish which had been thymectomized for 5 months before immunization. However, 9-month thymectomized fish had lower titres than controls. The response to AS was vigorous and significant memory was demonstrated after a second injection. There was no difference in response between control and thymectomized fish. The individual variation in response was not related to body size, antigen dose, sex or reproductive state. The results are discussed in terms of the thymus dependency or independency of antigens in fish, and the role and life span of T cell subsets, such as 'helper' and ';suppressor' cells.     

First report on antibody response of Seriola dumerilii (Risso 1810) challenged with Listonella anguillarum

The greater amberjack, Seriola dumerilii (Risso 1810) is a semi-pelagic fish and a worldwide species; it is considered a promising candidate for the diversification of Mediterranean aquaculture. In this paper an experimental injection with Listonella (Vibrio) anguillarum was performed to study the immune response of S. dumerilii. Antibody titres to L. anguillarum O1 were determined with indirect-ELISA at different times over a period of 42 days. Results showed that the antibody levels against L. anguillarum were significantly higher in the challenged fish compared to the control. They started developing since the 5th day reaching the highest peak on day 20 after injection, indicating a fast response of the immune system. The observed antibody titre was very high versus L. anguillarum if compared to other fish species.     

Evaluation of immune response to bovine rotavirus following oral and intraperitoneal inoculation in mice

With a view to use mice as an experimental model for studying immune response to bovine rotavirus (BRV), the kinetics of humoral and cellular immune responses to BRV in mice were evaluated by immunizing through intraperitoneal and oral route with UK strain of BRV. Following immunization with BRV, anti-rotavirus antibodies was developed in mice. The mean log antibody titres as measured by ELISA in mice immunized by intraperitoneal route were significantly higher than those immunized by oral route. Significant cellular immune response was observed in BRV-immunized mice on stimulation with BRV antigen, as measured by lymphocyte proliferation assay. The thymidine uptake by splenic and mesenteric lymph-node cells of intraperitoneally immunized mice on stimulation with BRV was 21328 +/- 1225 and 739 +/- 55 CPM, respectively. The splenic cells showed significantly higher stimulation (stimulation index 12.98) as compared to those of mesenteric cells (stimulation index 1.57). Foot pad inoculation test showed maximum virus-specific delayed type hypersensitivity reaction at 24 hr post-challenge following primary immunization and at 18 hr post-challenge following secondary immunization. The results indicate that BRV immunization by intraperitoneal route generates more efficient immune response in mice than by oral route and this route may be used for immune response studies involving BRV infection.     

Personality influences tetanus-specific antibody response in adult male rhesus macaques after removal from natal group and housing relocation

Previous research has suggested that personality is related to immune function in macaques. Using a prospective design, we examined whether variation in the personality dimension "Sociability" in adult male rhesus macaques (Macaca mulatta) was related to the in vivo secondary antibody response to a tetanus toxoid booster immunization following removal from natal groups and relocation to individual housing. We also explored whether the timing of the immunization following relocation had an impact on the immune response. Blood was sampled at the time of booster immunization, at 14 and 28 days post-immunization, and approximately 9 months post-immunization. Plasma was assayed for tetanus-specific IgG by enzyme-linked immunoassay (ELISA). There was no difference between High- and Low-Sociable animals in antibody levels at the time of the booster immunization. Multivariate analysis of variance (MANOVA) revealed that High-Sociable animals had a significantly higher antibody response following relocation and immunization compared to Low-Sociable animals. There was no effect of timing of the immunization on the immune response. The results confirm that personality factors can affect animals' immune responses, and that the dimension Sociability may be influential in a male's response to social separation and relocation.     

Immune response of mice of different inbred lines to Clostridium oedematiens anatoxin

Mice belonging to a number of inbred strains were immunized intradermally with Cl. oedematiens alpha-toxoid. The immunization was repeated 30 days later. On the 20th and the 30th days after the first injection and on the 10th day after the second one the antibody level against the toxoid was determined in the blood of mice by the passive hemagglutination test. The maximum response to the primary immunization was observed in the mice of the C3H strain, and the minimum one--in mice of the DBA/2 strain; the difference was more than 30-fold. The rest of the strains used in the test (A,CBA, BALB/c, AKR, CC57BR) displayed an intermediate level of the immune response. The differences reduced after the repeated immunization. The immune response to this antigen in mice is supposed to be genetically controlled.     

Effects of X-irradiation on the immune response of guinea pigs to Q fever vaccine

The immune response of guinea pigs to Q fever vaccine following 75 to 250 R (60 to 180 rads) of acute whole-body irradiations was investigated. Complement-fixing (CF) antibody titers and protection against febrile response to challenge with virulent Coxiella burnetii were studied. Exposures ranging from 75 to 250 R, 24 hours prior to inoculation, did not detectably alter the CF antibody response. Similar results were observed with 175 R delivered 48 or 72 hours before immunization. Protection against febrile response to challenge with 10(3) median fever doses of C. burnetii was seen in animals irradiated with 175 R, 24 or 72 hours before immunization. Significant protection was detectable at 14, 21, and 42 days after immunization in both irradiated and nonirradiated animals. Acute irradiation of the degree studied increases the mortality in normal animals infected 15 to 17 days later with virulent C. burnetii. The lethal effect could be prevented by use of Q fever vaccine.     

Divergent selection for antibody production in common carp (Cyprinus carpio L.) using gynogenesis

A base population (n = 101) of carp, consisting of a single hybrid cross, was immunized with the hapten-carrier complex DNP-KLH. to perform a divergent selection for antibody response. Measurement of the DNP-specific antibody response at 12 and 21 days postimmunization, allowed the classification of a low number of individual carp as early/high (10%) or late/low (13%) responders. Three individuals defined as early/high and three defined as late/low responding, were gynogenetically reproduced to obtain corresponding homozygous progenies within one generation only. Upon immunization with DNP-KLH, the antibody response was found to be significantly higher in the early/high responder homozygous offspring. Although the homozygosity of the offspring apparently caused a (s)lower antibody response (compared with the base population), the differences between the high and low responder offspring do indicate a genetic influence on the antibody response. The realized heritability (h2) for antibody production was estimated at 0.37 ± 0.36. The present study provides the basis for a divergent selection of homozygous inbred carp lines with a genetically controlled difference in antibody response. These inbred lines will allow us to investigate relationship(s) between immune responsiveness and resistance to infectious diseases in fish.     

Role of C3 in humoral immunity. Defective antibody production in C3-deficient dogs

Previous studies have shown that animals pharmacologically depleted of C3 have impaired antibody responses. However, such C depletion is neither complete nor sustained, and the C3 cleavage products generated by C3 depletion can both enhance and inhibit the immune response. To clarify the role of C3 in humoral immunity, the antibody response of dogs with genetically determined total deficiency of C3 (C3D) was examined. Serum IgG levels of the C3D animals were within the normal range, but were significantly lower than levels seen in normal controls or C3D heterozygotes. Specific antibody production was defective: the antibody titers of C3D dogs in response to primary intravenous immunization with two different T cell-dependent Ag (sheep E and bacteriophage phi X-174) were markedly reduced when compared to either normal controls or C3D heterozygotes. After secondary immunization with T-dependent Ag, the total antibody titers were normal, but the C3D dogs made proportionately more IgM and less IgG antibody than did either control group. After i.v. immunization with a T cell-independent Ag (DNP-Ficoll), the C3D dogs had reduced levels of IgM and IgG antibody after primary and secondary immunization. Neither i.m. immunization nor the use of a 20-fold increase in Ag dose i.v. could correct the defect seen in the antibody response of C3D dogs. The results herein demonstrate that C3 plays a critical role in the generation of a normal humoral immune response.     

Macrophage transformation of mononuclear cells following primary and secondary experimental immunization with measles vaccine

Cell-mediated and humoral immune response was studied in guinea pigs receiving two immunizations with live measles vaccine l-16 in doses of 1000 TCD50/0.5 ml at an interval of 45 days. The results of this study showed that the maximum level of the macrophagal transformation of mononuclears and the most intensive synthesis of antimeasles antibodies were observed on day 10 after booster immunization. The intensification of cell-mediated and humoral immune response was found to depend on the initial immunological background. The animals having had high values of cell-mediated response before booster immunization showed a decrease in these values, while an increase in antibody titers in such animals was transitory.     

Effectiveness of enteric immunization in the development of secretory immunoglobulin A response and the outcome of infection with respiratory syncytial virus.

Cotton rats were immunized via intranasal, intradermal, or enteric routes with respiratory syncytial virus (RSV) or a live recombinant vaccinia virus expressing the RSV F glycoprotein (vaccinia F). The animals were tested for the appearance of RSV-specific antibody responses in the serum, bronchoalveolar lavage, and nasal wash after immunization and for virus replication 4 days after intranasal challenge with RSV. RSV antibody response in the serum and respiratory tract was demonstrated in all immunization groups and was significantly increased after intranasal challenge with RSV. Immunoglobulin A (IgA) antibody response in bronchoalveolar lavage fluid after intranasal or enteric immunization was two- to threefold higher than that after intradermal immunization. Nasal-wash IgA antibody response was not significantly different among three immunization groups, although mean antibody titer was the highest in intranasal immunization group. Complete resistance to replication of RSV challenge was observed in the lungs of cotton rats immunized by the intranasal or enteric routes, whereas a low level of replication was detected in the lungs of rats immunized intradermally. Enteric or intradermal immunization conferred partial protection to the upper respiratory tract, but complete protection of the upper respiratory tract was observed in the intranasal immunization group. These observations suggest that while enteric immunization is quite effective in inducing antibody responses in the respiratory tract, the magnitude of antiviral immunity induced in the respiratory tract after intranasal immunization may be superior to that observed after enteric immunization.     

卵形鲳鯵对刺激隐核虫的免疫应答和免疫保护研究

用刺激隐核虫(Cryptocaryon irritans)的幼虫对卵形鲳鯵(Trachinotus ovatus)进行腹腔注射和体表感染,然后每隔一周用阻动试验(Immobilization assay)检测免疫鱼的抗血清和皮肤培养液对刺激隐核虫幼虫的阻动效价,在第14周中,分别用亚致死剂量和致死剂量的刺激隐核虫幼虫对免疫鱼攻毒以检测所产生的免疫保护力。实验结果显示:两种免疫方法都能让卵形鲳鯵的血清和皮肤生成阻动刺激隐核虫幼虫的特异性抗体,并能使被免疫鱼获得明显的免疫保护,但是体表感染免疫组的血清和皮肤培养液的阻动效价都要比腹腔注射免疫组高,所获得的免疫保护力也更强。同时还发现,免疫鱼血清和皮肤培养液中的抗体存在明显的差异:两者的最初生成时间、达到峰值的时间、变化规律以及阻动效价等都不一致。因此,我们推测鱼类的系统免疫应答和皮肤黏膜免疫应答有可能是相互独立的,或者是不同步的。鱼类的体液免疫应答,特别是黏膜免疫应答对抵御刺激隐核虫的感染起了重要的作用,采用刺激隐核虫虫体疫苗可能成为预防海水鱼类白点病的一种选择。