Serum antibody does not account for differences in the severity of chronic respiratory disease caused by Mycoplasma pulmonis in LEW and F344 rats

Chronic respiratory disease in rats, resulting from Mycoplasma pulmonis infection, is useful in the study of the immunological mechanisms in similar inflammatory diseases and provides a unique opportunity to study the interactions between systemic and mucosal immune systems in a naturally occurring infection. The present study examined the serum antibody responses to M. pulmonis in strains of rats which differ in disease progression and severity; LEW rats developed more severe disease than did F344 rats. Serum antibody responses were evaluated as to their levels, isotypes, and antigens recognized. Infected LEW rats produced greater or equal levels of the major classes of serum antibody to M. pulmonis than did infected F344 rats, suggesting that development of serum antibody responses alone does not resolve lesions and is not responsible for the difference in disease severity found in LEW and F344 rats. Although LEW rats produced higher responses in all subclasses of immunoglobulin G (IgG), the specific IgG response of LEW rats was composed predominately of IgG1 and IgG2a subclasses, while IgG2b was the major component of the IgG response in F344 rats. Finally, LEW rats responded more quickly to M. pulmonis antigens than did F344 rats, and there was no difference in the antigens eventually recognized by each strain, confirming previous work which suggested that LEW rats do not exhibit an unresponsiveness to a specific antigen(s) of M. pulmonis.     

Environmental modulation of lipopolysaccharide chain length alters the sensitivity of Escherichia coli to the neutrophil bactericidal/permeability-increasing protein.

The plaque-forming cell (PFC) assay with sheep erythrocytes (SRBC) sensitized with different antigens and a 4-h tritiated thymidine pulse assay were used to determine whether polyclonal activation occurs in rats following in vivo administration of Mycoplasma pulmonis. Injection of M. pulmonis into F344 rats resulted in an increase in the number of splenic immunoglobulin M-secreting PFC that produced antibodies reactive with the trinitrophenyl hapten and with SRBC. This polyclonal response reached a peak by 72 h after injection and returned to normal levels by 96 h, at which time the specific response to M. pulmonis reached its peak. Heat treatment and preopsonization of M. pulmonis with antiserum before injection resulted in reduced numbers of PFC against M. pulmonis-sensitized SRBC, trinitrophenyl hapten-sensitized SRBC, and SRBC. The number of PFC against the three types of target cells also increased in LEW rats after immunization with M. pulmonis. The number of PFC against SRBC and staphylococcal protein A-sensitized SRBC was higher in immunized LEW rats than in immunized F344 rats. Examination of unimmunized animals also revealed that LEW rats had higher initial numbers of PFC than did F344 rats. These results showed that polyclonal activation occurs in rats following in vivo administration of M. pulmonis and that LEW rats have an inherent propensity to develop higher nonspecific responses in vivo than F344 rats.     

Murine respiratory mycoplasmosis in F344 and LEW rats: evolution of lesions and lung lymphoid cell populations.

By comparison of two strains, LEW and F344, which are known to differ in susceptibility to Mycoplasma pulmonis respiratory disease, it was shown that differences in lesion severity and progression were associated with changes in lung lymphocyte populations. Lung lesions in LEW rats developed earlier after infection, became more severe, and were characterized by continued proliferation of all classes of lymphoid cells, T lymphocytes, B lymphocytes, and plasma cells, throughout the 120-day observation period. In contrast, lymphoid proliferation in F344 rats reached a plateau at 28 days and was restricted to an increase in T lymphocytes, immunoglobulin A (IgA)-bearing B lymphocytes, and IgA and IgG plasma cells. Although approximately 10 times as many IgG B cells and 4 times as many IgG plasma cells were found in infected LEW rats as compared with F344 rats, the specific anti-M. pulmonis IgG response in the two strains was roughly parallel. The same relationships held true, although to a lesser extent, for specific IgA antibody responses and cellular responses. Whereas lung lesions showed a tendency to resolve in F344 rats by 120 days, severe lesions persisted in LEW rats. The disparity between the cellular response and specific antibody response, the seemingly uncontrolled lymphocyte proliferation in LEW rats, and the mitogenic potential of M. pulmonis suggest that differences between LEW and F344 rats in lung lesion severity and progression are related to differences in the degree of nonspecific lymphocyte activation in the two strains, an imbalance in regulation of lymphocyte proliferation in LEW rats, or both.     

Specific and nonspecific antibody responses in different segments of the respiratory tract in rats infected with Mycoplasma pulmonis.

Murine respiratory mycoplasmosis resulting from Mycoplasma pulmonis infection in rats provides a useful model for the study of immunological and inflammatory mechanisms operative in the respiratory tract. We have previously shown that LEW rats develop more severe disease than do F344 rats. To further study the production of antibody responses in chronic respiratory disease due to M. pulmonis infection, we examined the distribution and development of M. pulmonis-specific antibody-forming cells (AFC) in different segments of the respiratory tracts of infected LEW and F344 rats. In these studies, the upper respiratory nodes were the initial site of antibody production after infection and remained the major site for recovery of AFC. Since infected LEW rats had equal or higher numbers of AFC than did infected F344 rats, these results suggest that the level of local antibody production alone is not responsible for the decreased susceptibility of F344 rats to murine respiratory mycoplasmosis. The differences in total antibody responses appear to be due to the greater numbers of cells recovered from the tissues of infected LEW rats compared with those recovered from F344 rats, suggesting that LEW rats may have greater production of chemotactic factors. Also, we demonstrate that nonspecific activation and/or recruitment of B cells occurs in the respiratory tracts of both LEW and F344 rats after infection with M. pulmonis.     

Nonspecific lymphocyte responses in F344 and LEW rats: susceptibility to murine respiratory mycoplasmosis and examination of cellular basis for strain differences.

Mycoplasma pulmonis produces a mitogen which may play a role in the pathogenesis of murine respiratory mycoplasmosis in rats. Since LEW rats are more susceptible to this disease than F344 rats are, these two strains were used to examine a possible association between disease severity and the level of nonspecific lymphocyte stimulation by mitogens, including M. pulmonis membrane preparations. F344 and LEW spleen, lung, blood, and lymph node lymphocytes were exposed to various mitogens. LEW lymphocytes gave a significantly higher response to mitogenic stimulation, regardless of their anatomical source. These differences in lymphocyte responsiveness were primarily due to differences within the nonadherent cell population. Significantly higher numbers of W3/25+ (T helper) cells were found in LEW lymphoid populations, whereas no difference was found in MRC OX-8+ (T suppressor/cytotoxic) cells. These data suggest an association between disease severity and host responsiveness to nonspecific stimuli.     

Antibody response against perlecan and collagen types IV and VI in chronic renal allograft rejection in the rat

Chronic rejection is the leading cause of late renal transplant failure. Various structural lesions are observed in grafts undergoing chronic rejection including glomerular basement membrane (GBM) duplications. The well-established Fisher (F344) to Lewis (LEW) rat renal transplant model for chronic rejection was used to assess the presence and role of the humoral immune response against graft antigens during chronic rejection. LEW recipients of F344 allografts develop transplant glomerulopathy and produce IgG1 antibodies directed against F344 GBM preparations that are detectable 3 weeks after transplantation. Glomerular IgG1 deposition was observed that in vitro co-localized with a rabbit anti-rat GBM antiserum in rejecting F344 grafts; elution experiments of isolated glomeruli yielded IgG1 antibodies reactive in vitro with F344 GBM, but not LEW GBM. Prevention of acute rejection by transient treatment of the recipients with cyclosporin A completely abrogated the production of anti-GBM antibodies. Using proteomic techniques we identified the antigens recognized by the LEW posttransplant sera as being the heparan sulfate proteoglycan perlecan and the alpha1 chain of collagen type VI in association with the alpha5 chain of collagen type IV. In conclusion, LEW recipients of F344 kidney grafts produce IgG1 antibodies against donor type perlecan and alpha1(VI)/alpha5(IV) collagen and develop transplant glomerulopathy. These data implicate an important role for the humoral immune response in the development of glomerulopathy during chronic rejection.     

Immunoglobulin class- and subclass-specific responses to Mycoplasma pulmonis in sera and secretions of naturally infected Sprague-Dawley female rats

Mycoplasma pulmonis causes chronic murine respiratory mycoplasmosis and genital disease in rats. Specific immunoglobulin M (IgM), IgA, and IgG and its subclasses present in sera and tracheal and uterine lavage samples from 36 naturally infected Sprague-Dawley female rats were tested for reactivity with M. pulmonis in an enzyme-linked immunosorbent assay. Ten specific-pathogen-free Sprague-Dawley female rats served as the negative controls. Tracheal and uterine lavage samples were cultured quantitatively for M. pulmonis. M. pulmonis was isolated from the trachea (35 of 36) and uterus (17 of 36) of naturally infected rats; all rats were infected in at least one of the two sites cultured. M. pulmonis was not isolated from any control rat. There was a significant difference in levels of specific antibody of all classes except IgG2c between control and naturally infected animals (P less than 0.001 for IgM, IgG, IgG1, and IgG2a; P less than 0.002 for IgG2b; and P less than 0.02 for IgA). There was no correlation between numbers of M. pulmonis cells isolated and the amount or class of antibody measured in serum or tracheal lavage specimens. The predominant antibodies to M. pulmonis found in the sera of naturally infected rats were IgG and IgM. The IgG2a subclass was responsible for the majority of IgG-positive animals. There were no differences between rats which were positive by culture for M. pulmonis in the uterus (U+) and rats which were negative by culture for M. pulmonis in the uterus (U-) with respect to distribution or amount of antibody classes and subclasses in the serum. However, tracheal wash samples from U+ rats had significantly higher (P less than 0.03) levels of specific IgG1 and IgG2a than those from U- rats. Conversely, IgG2a was present in higher levels in pooled uterine lavage specimens from U- rats than in those from U+ rats.     

CD26 (dipeptidyl-peptidase IV)-dependent recruitment of T cells in a rat asthma model

CD26 truncates several chemokines as well as neuropeptides and influences immune responses via modulation of cell adhesion and T cell activation, suggesting an involvement of CD26 in asthmatic and airway inflammation. Therefore, Fischer 344 (F344), Brown Norway (BN) and Lewis (LEW) rat strains, which differ in their CD26-like enzymatic activity, were compared using an asthma model. Additionally, two CD26-deficient mutant F344 rat substrains were included and compared to the wild-type F344 substrain. Immunization was performed twice with ovalbumin (OVA), and 2 weeks later the rats were challenged with OVA intratracheally Flow cytometry (FACS) analysis of different leucocyte subsets as well as enzyme-linked immunosorbent assay (ELISA) for IgE levels in the blood and bronchoalveolar lavage (BAL) were performed 24 h after challenge. LEW rats with the lowest CD26 activity among the rat strains investigated here displayed significantly reduced CD4+ T cell numbers in the BAL compared to wild-type F344 and BN rats. Moreover, in asthma, the ratio of CD26+ to CD26- T cell receptor (TCR)-positive cells increased significantly in F344 and LEW but not BN rats. Most intriguingly, in both CD26-deficient F344 rat substrains the number of CD4+ T lymphocytes was markedly reduced compared to wild-type F344. The decrease in T cell recruitment observed in the CD26-deficient rats was associated with significantly reduced OVA-specific IgE-titres. This is the first report to show a remarkably reduced T cell recruitment in rat strains that either lack or exhibit reduced CD26-like enzymatic activity, suggesting a role for CD26 in the pathogenesis of asthma via T cell-dependent processes such as antibody production.     

Effects of aging on exocrine immune responses to Mycoplasma pulmonis.

Formalinized Mycoplasma pulmonis was used to immunize 3 different age groups of Fischer 344 rats. A specific antibody to this antigen was detected in both saliva and lung lavage fluids and differences were noted in the elicitation of secretory antibody between the different ages of the animals. Few statistical differences were noted between the three age groups for salivary IgG responses to M. pulmonis, regardless of the dosage given, even though all responses were greater than their respective control groups. The principal differences among the three age groups were noted in the kinetics of the response, that is, the amount of time that was necessary to produce a peak response. The younger group of animals took less time to produce a peak response than the older two groups, even though the magnitude of the response was lower. Salivary IgA responses to M. pulmonis appeared predominantly as a primary response, particularly in the senescent animals. Secondary salivary IgA responses were not significantly greater than their respective primary responses, suggesting that secretory IgA did not display classic anamnestic responses that were observed with salivary IgG. As with IgG responses, the senescent animals took longer to produce a peak salivary IgA response when compared to the other age groups. Lung lavage IgG responses, normalized to total protein, were greatest in the youngest group of animals and appeared to diminish as the age of the animal increased. In contrast, lung lavage IgA responses to M. pulmonis were of a greater magnitude in the senescent animals. These studies suggest that senescent animals are capable of eliciting a humoral immune response in mucosal secretions to Mycoplasma pulmonis. However, differences noted with regard to disease severity and mortality to respiratory mycoplasmosis in senescent animals may result from intrinsic defects in the quality of the humoral response or as a consequence of deficient cellular responses to this pathogen.     

Effects of 3,4-methylenedioxymethamphetamine on locomotor activity and extracellular dopamine in the nucleus accumbens of Fischer 344 and Lewis rats

Previous studies have shown that Fischer 344 (F344) and Lewis (LEW) rats may differ with respect to their behavioural and neurochemical responses to several drugs of abuse, including amphetamines. Herein, we have examined whether such strain differences extend to a ring-substituted amphetamine, namely 3,4-methylenedioxymethamphetamine (MDMA, ecstasy), a recreationally-used drug endowed with euphoric, but also long-term neurotoxic effects. Beside strain differences in baseline locomotor activity (F344>LEW), it was found that the subcutaneous administration of 10 mg/kg, but not 5 mg/kg, MDMA increased locomotor activity in F344 rats only. On the other hand, such a treatment increased to similar extents extracellular dopamine (DA) levels in the nucleus accumbens of F344 and LEW rats, thus suggesting that genetic differences in MDMA locomotor effects are not accounted for by accumbal DA release.     

Regulation of experimental autoimmune uveitis in rats--separation of MHC and non-MHC gene effects.

Experimental autoimmune uveitis (EAU) is an organ-specific autoimmune disease and has served as a model of certain ocular inflammatory conditions in man. The present study was aimed at separating the effects of MHC and non-MHC genes on the development of EAU in the rat. EAU-susceptible LEW (RT1l), EAU-resistant WKAH (RT1k), and WKAH.1L (RT1l) MHC congenic strain of WKAH background rats were immunized with retinal soluble antigen (S-Ag) in Freund's complete adjuvant (FCA). LEW rats showed typical EAU, while neither WKAH nor WKAH.1L congenic rats developed EAU. However, when an additional i.v. injection of Bordetella pertussis was given, all rat strains developed EAU. Furthermore, when immunized with peptide M, an 18-mer synthetic peptide, which corresponds to amino acid positions 303-320 of bovine S-Ag, and given an additional i.v. injection of B. pertussis, LEW and WKAH.1L rats developed EAU, whereas WKAH did not. When ACI (RT1avl), BUF (RT1b), LEJ (RT1j), W (RT1k), F344 (RT1lvl), BN (RT1n), NIG-III (RT1q), TO (RT1t), and SDJ (RT1u) rats were immunized with peptide M or S-Ag and then B. pertussis, all strains developed EAU by immunization with S-Ag plus B. pertussis, but only F344 and NIG-III developed EAU by immunization with peptide M. These findings suggest that susceptibility to EAU in rats is controlled by both MHC and non-MHC genes; and that in the absence of B. pertussis adjuvant, the form of disease induced by native S-Ag in FCA is governed by non-MHC gene(s). However, this effect of non-MHC gene(s) could no longer be observed when the rats were also injected with B. pertussis adjuvant at sensitization.     

Effects of Cross Fostering on Open-Field Behavior, Acoustic Startle, Lipopolysaccharide-Induced Corticosterone Release, and Body Weight in Lewis and Fischer Rats

Lewis (LEW/N) and Fischer (F344/N) rats differ on a myriad of behavioral and physiological endpoints, some of which have been reported to be affected by maternal experience in outbred rats and other strains. To assess whether epigenetic factors contribute to the differential behavioral responses to stress and pro-inflammatory challenges in these strains, the effects of cross fostering on open-field, acoustic startle, and glucocorticoid reactivity to lipopolysaccharide (LPS) were examined in the present experiment. In the open-field test, although in-fostered female LEW/N and F344/N strains did not differ, female LEW/N rats displayed significantly greater activity than female F344/N rats in the cross-fostered condition. Differences between males of the two strains were increased by cross fostering, with the LEW/N strain displaying greater total activity. In acoustic startle, there was little strain difference between in-fostered or cross-fostered female rats. On the other hand, in-fostered male LEW/N rats had a significantly greater startle response than in-fostered male F344/N rats, an effect that was dramatically reduced by cross fostering. In-fostered female LEW/N rats displayed a blunted corticosterone response relative to in-fostered female F344/N rats, an effect that was reduced by cross fostering. Conversely, although there was no strain difference between male in-fostered rats, cross-fostered male F344/N rats displayed a significantly greater corticosterone response to LPS than cross-fostered male LEW/N rats. Finally, body weight differences between in-fostered LEW/N and F344/N rats were reduced by cross fostering. Together, these data illustrate that maternal factors play a role in the behavioral and physiological responses to stress between the two strains.     

Hapten antibody production and the relevance of allogeneic reactions to elimination of the carrier effect

Lewis (LEW) rats immunized 3 weeks before by injection of DNP-KLH together with Bordetella pertussis showed high levels of DNP antibody as judged by serum binding of 10(-7) M 3H-DNP-lysine 10 days after secondary immunization with DNP-KLH. Sera obtained from LEW rats following secondary immunization with DNP-BGG showed reduced DNP hapten binding. However, injection of 10(8) F1 hybrid Lewis X Brown Norway spleen cells into DNP-primed LEW rats 2 days before secondary immunization with DNP-BGG significantly increased the level of serum binding of 3H-DNP-lysine. These results provide evidence that the allogeneic cellular reaction associated with a host-versus-graft response induced by injection of F1 hybrid lymphoid cells into DNP-primed parental strain recipients partially obviates the requirement for carrier-specific T cells in the secondary anti-DNP response thereby providing a stimulus for triggering primed host B cells to produce antibody.     

The avidity of IgM antibody in high and low responder rats.

This study examined IgM antibody produced by highly responding ACI and poorly responding F344 rats follwing immunization with poly(Glu52Lys33Tyr15) or poly(Glu52Lys33Tyr15) aggregated with methylated bovine serum albunim (MeBSA). The ACI rats produced both IgM and IgG plaque-forming cells (PFC) following immunization with either form of antigen. The F344 rats did not respond to unaggregated poly(Glu52Lys33Tyr15), but they produced significant amounts of IgG PFC and extremely small amounts of IgM PFC after immunization with poly(Glu52Lys33Tyr15)/MeBSA. Both high and low responder rats had similar kinetic profiles of IgM antibody production, and this antibody had nearly identical avidity in both strains with no evidence for any maturation in avidity. thus, one of the genetic defects in the antibody response to poly(Glu52Lys33Tyr15) is an inability of the F344 strain to produce large amounts of IgM in response to this antigen.     

Rat strains differ in susceptibility to Ureaplasma parvum-induced urinary tract infection and struvite stone formation

Individuals with struvite uroliths are susceptible to recurrent urinary tract infections (UTI), sepsis, and renal disease. Unfortunately, little is known about the host-specific factors that predispose to this disease. In order to develop a rodent model that can address this problem, we inoculated female Fischer 344 (F344), Lewis (LEW), Sprague-Dawley (SD), and Wistar (WIS) rats with a host-adapted strain of Ureaplasma parvum. Animals were necropsied at 2 weeks postinoculation; 100% of F344, 42% of SD, 10% of LEW, and 10% of WIS rats remained infected. Severe bladder lesions and struvite calculi were seen in 64% of F344 rats; in other rat strains, bladder lesions were mild or absent. F344 rats with struvite uroliths had the highest urinary levels of proinflammatory cytokines, such as GRO/KC, interleukin-1alpha (IL-1alpha), and IL-1beta. F344 rats without struvite stones at necropsy had milder bladder lesions and significantly lower urinary levels of proinflammatory cytokines but a more prominent inflammatory response than did other rat strains. Based on our results, struvite stone formation is linked to a robust inflammatory response that does not resolve UTI but instead promotes damage to surrounding tissues.     

Rat strain differences in freezing and sleep alterations associated with contextual fear

STUDY OBJECTIVE: To examine rat strain differences in fear responses and subsequent alterations in sleep associated with contextual fear. DESIGN: Recordings for each strain were obtained of nondisturbed baseline sleep and of sleep after exposure to a novel chamber (handling control). Afterward, the rats were subjected to shock training for 2 sessions (ST1, ST2) and to contextual fear (CF) alone. Percentage time spent in freezing (FT%) was observed during ST1, ST2 and CF exposures. Sleep was recorded for 20 hours (8-hour light and 12-hour dark period) following ST1, ST2 and CF. SETTING: NA SUBJECTS: The subjects were 2 inbred rat strains (Fischer 344 [F344] and Lewis [LEW] and one outbred rat strain (Wistar [WST]). INTERVENTIONS: The rats were surgically implanted with electrodes for recording electroencephalogram and electromyogram for determining arousal state. MEASUREMENTS AND RESULTS: Strain rankings for FT% were F344 = LEW > WST during ST2 and CF. LEW and WST rats exhibited decreased rapid eye movement sleep (REM) after shock training and CF compared with baseline and control; F344 rats did not. F344 and WST rats showed increased dark-period REM after ST1, ST2, and CF compared with baseline but not with control. CONCLUSIONS: Shock training and CF induce immediate reductions in REM in rats. However, strain differences in the amount of REM decrease did not simply relate to the strength of fear responses during shock training and CF. We speculate that reported strain differences in stress hormones, particularly prolactin, may contribute to strain differences in fear-induced alterations in REM.     

Intracage ammonia promotes growth of Mycoplasma pulmonis in the respiratory tract of rats.

Ammonia (NH3) from soiled cage bedding is known to enhance the progression and severity of murine respiratory mycoplasmosis in rats. To test the hypothesis that NH3 directly or indirectly enhances the growth of Mycoplasma pulmonis in vivo, pathogen-free F344 rats were inoculated intranasally with 1 x 10(4) to 4 x 10(4) or 4 x 10(6) to 5 x 10(6) colony-forming units of M. pulmonis and exposed to less than or equal to 1.5 or 76 microgram of NH3 per liter (less than or equal to 2 or 100 ppm, respectively). Nasal passages, larynges, tracheas, and lungs from rats killed at intervals up to 28 days after inoculation were quantitatively cultured. Growth of M. pulmonis was much greater in NH3-exposed rats than in controls, particularly in those inoculated with the lower dose. Increases in M. pulmonis populations were more rapid in proximal airways than in distal airways. Serum immunoglobulin G and M antibody responses to M. pulmonis as measured by an enzyme-linked immunosorbent assay were greater in NH3-exposed rats. In other experiments, the nasal passages absorbed virtually all NH3 when the rats were exposed to less than 380 micrograms of NH3 per liter (500 ppm), indicating that NH3 induced increases in the numbers of organisms in the distal respiratory tract, probably by a secondary, rather than a direct, effect. Also, NH3 exposure did not inhibit pulmonary antibacterial activity as measured by clearance of radiolabeled Staphylococcus epidermidis. The growth of M. pulmonis in vitro was inhibited by 1 mM NH4+ added to the medium as NH4OH but not by NH4+ concentrations of 0.5, 0.1, or 0.01 mM, suggesting that NH3 increases growth indirectly through effects on the host.     

Systemic and Mucosal Immune Responses after Intranasal Administration of Recombinant Mycobacterium bovis Bacillus Calmette-Guérin Expressing Glutathione S-Transferase from Schistosoma haematobium

A major goal of current vaccine development is the induction of strong immune responses against protective antigens delivered by mucosal routes. One of the most promising approaches in that respect relies on the use of live recombinant vaccine carriers. In this study, Mycobacterium bovis BCG was engineered to produce an intracellular glutathione S-transferase from Schistosoma haematobium (Sh28GST). The gene encoding Sh28GST was placed under the control of the mycobacterial hsp60 promoter on a replicative shuttle plasmid containing a mercury resistance operon as the only selectable marker. The recombinant Sh28GST produced in BCG bound glutathione and expressed enzymatic activity, indicating that its active site was properly folded. Both intraperitoneal and intranasal immunizations of BALB/c mice with the recombinant BCG resulted in strong anti-Sh28GST antibody responses, which were enhanced by a boost. Mice immunized intranasally produced a mixed response with the production of Sh28GST-specific immunoglobulin G1 (IgG1), IgG2a, IgG2b, and IgA in the serum. In addition, high levels of anti-Sh28GST IgA were also found in the bronchoalveolar lavage fluids, demonstrating that intranasal delivery of the recombinant BCG was able to induce long-lasting secretory and systemic immune responses to antigens expressed intracellularly. Surprisingly, intranasal immunization with the BCG producing the Sh28GST induced a much stronger specific humoral response than intranasal immunization with BCG producing the glutathione S-transferase from Schistosoma mansoni, although the two antigens have over 90% identity. This difference was not observed after intraperitoneal administration.     

Direct identification of rat iNKT cells reveals remarkable similarities to human iNKT cells and a profound deficiency in LEW rats

iNKT cells are a particular lymphocyte population with potent immunomodulatory capa‐city; by promoting or suppressing immune responses against infections, tumors, and autoimmunity, iNKT cells are a promising target for immunotherapy. The hallmark of iNKT cells is the expression of a semiinvariant TCR (with an invariant α‐chain comprising AV14 and AJ18 gene segments), which recognizes glycolipids presented by CD1d. Here, we identified iNKT cells for the first time in the rat using rat CD1d‐dimers and PLZF staining. Importantly, in terms of frequencies (1.05% ± 0.52 SD of all intrahepatic αβ T cells), coreceptor expression and in vitro expansion features, iNKT cells from F344 inbred rats more closely resemble human iNKT cells than their mouse counterparts. In contrast, in LEW inbred rats, which are often used as models for organ‐specific autoimmune diseases, iNKT cell numbers are near or below the detection limit. Interestingly, the usage of members of the rat AV14 gene family differed between F344 and LEW inbred rats. In conclusion, the similarities between F344 rat and human iNKT cells and the nearly absent iNKT cells in LEW rats make the rat a promising animal model for the study of iNKT cell‐based therapies and of iNKT‐cell biology.     

A Study on the Glycan Specificity of Natural Antibody Repertoires in Rodents

Inbred strains of mice and rats are widely used in preclinical investigations evaluating the effectiveness of glycan-based biomecines, however, the glycan specificity repertoires of serum Abs in rodents have not been fully characterized. In the present study, serum antibodies in naïve mice and rats of different inbred strains were analyzed for specificity against 4 representative carbohydrate structures including PGA (1,4-linked α-D-galactopyranosyluronic acids), β-glucan, mannan and α-glucan (dextran). Mannan was not recognized by serum Abs from any of the mouse and rat strains. Serum IgM in naïve F344, BN and Lewis rats recognized PGA and β-glucan and, less strongly, dextran. High titer circulating IgM against PGA were found in mice of BALB/c, C57BL/6, C3H/NeH and BXSB strains. C3H/NeH was the only strain which also produced low titer IgM against β-glucan and dextran. Age-related production of high titer IgM, IgA and IgG Abs against β-glucan was observed in BXSB mice. Intraperitoneal immunization of BALB/c and C57BL/6 mice with β-glucan elicited strong IgM responses, while immunization with PGA also led to an increase of anti-PGA IgM Ab titers. These results provide useful information on the characteristics of glycan-specific natural antibody repertoires in rodents.