The effect of anti-lymphocytic antibody on the humoral immune response in different strains of mice: I. The response to bovine serum albumin

The effect of a single batch of horse anti-mouse thymocyte globulin on the humoral immune response has been extensively investigated in 10–12-week-old male mice of six inbred strains. The preparation under test readily suppressed the primary immune response of A/HeJ, C57B1, DBA/1, CBA and C₃H mice to 0.1, 1.0 and 10.0 mg doses of alum precipitated bovine serum albumin (alum BSA). However, at the doses used it generally failed to significantly suppress the primary immune response of BALB/c mice to 0.1 and 1.0 mg doses of this antigen. Furthermore, the effectiveness of the suppression was somewhat diminished by the incorporation of bordetella pertussis organisms into the alum BSA inoculum. The product also suppressed the secondary immune response to alum BSA in C₃H mice and C57B1 mice but, at the same doses, was without significant effect on the secondary responses of A/HeJ and CBA mice.     

The glucuronoxylomannan of Cryptococcus neoformans serotype A is a type 2 T-cell-independent antigen.

The humoral immune response of inbred mice to immunization with the glucuronoxylomannan (GXM) of Cryptococcus neoformans was investigated both serologically and in plaque-forming cells (PFCs). The T-helper-cell-independent quality of the GXM was demonstrated by using BALB/c nu/nu mice. Primary and secondary dose responses to three antigenic forms of GXM, (i) the native antigen, (ii) a GXM-bovine serum albumin protein conjugate, and (iii) a cryptococcal whole-cell vaccine, revealed a lack of isotype class switching and anamnestic responses. Both the levels of complement-fixing anti-GXM antibody in serum and the PFC responses in the athymic mice showed no significant differences from those in the wild-type controls. However, T cells are involved in the suppression of the primary response to GXM. When BALB/cBy mice were given rabbit anti-mouse thymocyte serum along with 0.5 microgram of GXM, both antibody levels in serum and PFC responses were significantly increased over those of control mice that received GXM and normal rabbit serum. In addition, T cells were also shown to enhance the primary immune response to GXM. BALB/cBy mice were given GXM and anti-mouse thymocyte serum on day 1. On day 2, the experimental group was given anti-mouse thymocyte serum and the control group was given saline. On day 5, comparison of the PFC responses and anti-GXM antibody titers of the two groups revealed a significant increase in the immune response of the control over the experimental group. The type 2 T-cell-independent quality of GXM was also demonstrated in CBA/cHN xid mice. These mice lack the Lyb+ subset of B cells and are unable to respond to type 2 T-independent antigens but respond normally to type 1 T-independent antigens. Type III pneumococcal polysaccharide, a type 2 T-independent antigen, was used as a negative control, and trinitrophenyl-lipopolysaccharide, a type 1 T-independent antigen, was used as a positive control. The CBA/cHN xid mice failed to respond to either type III pneumococcal polysaccharide or GXM but did not respond to immunization with trinitrophenyl-lipopolysaccharide. BALB/cBy mice responded normally to all three antigens.     

The effect of antilymphocytic antibody on the humoral immune response in different strains of mice: II. The response to sheep erythrocytes

The effect of an horse anti-mouse thymocyte globulin (ALG) preparation on the primary and secondary immune responses to sheep erythrocytes (RBC) was studied in 2–3-month-old male A/HeJ, C57B1, BALB/c, DBA/1, CBA and C3H mice. A wide variation in the primary and secondary responses of the individual strains was noticed but in all cases these were significantly suppressed by ALG pretreatment. The capacity of the ALG to suppress the primary immune response was not overcome by increasing the antigen dose from 3 × 10⁷ to 3 × 10⁹ sheep RBC.     

Modulation of the immune response to pneumococcal type 14 capsular polysaccharide-protein conjugates by the adjuvant Quil A depends on the properties of the conjugates

Streptococcus pneumoniae type 14 capsular polysaccharide-bovine serum albumin (S14PS-BSA) conjugates were prepared by water-soluble-carbodiimide-mediated condensation with or without the use of N-hydroxy-sulfosuccinimide. The immunogenicities of the capsular polysaccharide (S14PS) and of the conjugates were studied in (CBA/N x BALB/c)F1 mice and in female BALB/c mice. The response in these mice indicates that S14PS could be classified as a thymus-independent type 2 antigen. Coupling of S14PS to BSA improved the immunogenicity of this polysaccharide, and an immunoglobulin G memory response was evoked. Conjugation with N-hydroxysulfosuccinimide resulted in a product with a higher polysaccharide/protein ratio. This conjugate induced a greater immune response than did the classical conjugate. Quil A enhanced the immune response to S14PS and to most S14PS-BSA conjugates. The enhancement of the immune response to the conjugates seemed to depend on the coupling procedure. Our results indicate that for the construction of immunostimulating complexes based on polysaccharide or oligosaccharide-protein conjugates, attention should be paid to the degree of cross-linking of the antigens involved.     

Studies on immunological paralysis. II. The detection and significance of antibody-forming cells in the spleen during immunological paralysis with type III pneumococcal polysaccharide

The immunocytoadherence (rosette) technique has been adapted to study the immune response to varying doses of type III pneumococcal polysaccharide (SIII). Syngeneic erythrocytes coated with SIII were incubated with washed spleen cell suspensions isolated 0·5–39 days after i.v. injection into CBA strain mice of immunogenic (0·5 or 5·0 μg) or paralysing (500 μg) doses of SIII; the immunizing or paralysing effect of these doses of SIII was confirmed by a passive haemagglutination technique. The number of rosette-forming cells per spleen was significantly increased above the background range in twenty-seven out of forty-two, twenty-four out of thirty-seven and forty-one out of forty-six animals injected with 0·5, 5·0 and 500 μg SIII, respectively. Not only was there a greater incidence of `positive' values in the paralysed group, but the response was better maintained than in the immunized groups.

In view of the apparent lack of central inhibition in these experiments, paralysis with SIII was considered to be attributable to the antibody-neutralizing effect of persisting undegraded antigen. It is suggested that the phenomenon of polysaccharide paralysis may be the expression of more than one mechanism.

     

The effect of antilymphocytic antibody on the humoral immune response of different strains of mice: IV. Variable effect of individual ALG preparations on the immune response to type III polysaccharide antigen

ALG has been isolated from the sera of four horses immunized with mouse thymocytes and its ability to suppress the immune response to SSS-III investigated. All four preparations suppressed the response to this antigen in BALB/c mice but only one was effective in CBA mice. Other ALGs were isolated from sequential bleeds obtained from a single horse. Preparations obtained after only two injections of the donor with mouse thymocytes, were able to suppress the immune response to SSS-III in CBA and BALB/c mice. Studies with selected ALG preparations indicated that they were less effective or ineffective if their administration was delayed until just before challenge with SSS-III. These results indicate that the ability of ALG to suppress thymus-independent immune processes is a variable one, but it is not necessarily a consequence of hyperimmunization of the donor animal. It is however influenced by the ALG protocol used and the strain of animal under test.     

Streptococcal group A carbohydrate antibodies in mice: evidence for strain differences in magnitude and restriction of the response, and for thymus dependence

Immunization with streptococcal group A vaccines revealed significantly higher immune responses in BALB/c than C57BL/6, CBA, and DBA/2 mice. In general, the antibody response of BALB/c mice not only was markedly higher than that of the other strains, but also showed a high degree of restriction. With two exceptions, all immune sera of the BALB/c mice exhibited bands in very similar electrophoretic positions. On the other hand, the three other strains were low responders which, except for the DBA/2 mice, rarely demonstrated clear-cut bands in the immune sera. Studies with thymusless (nude) mice carrying 50 – 85% BALB/c genome identified the streptococcal group A polysaccharide, presented on whole bacteria, as a thymus-dependent antigen. These nude mice, after reconstitution with thymus cells, produced group A antibodies restricted to two antibody bands extremely similar to those of BALB/c mice. High levels of antibody were produced in mice reconstituted with low responder DBA/2 thymus cells, indicating that the magnitude of the response is entirely controlled by mechanisms at the bone marrow cell level. Magnitude and restriction of the antibody response to this antigen are not determined by the H-2 allele and are not related to the Ig^a allotypic marker on the heavy chain.     

Dynamics of immune response to porine fromYersinia pseudotuberculosis outer membrane

The dynamics of immune response to pore-forming protein isolated from the outer membrane ofYersinia pseudotuberculosis was studied on CBA and BALB/c mice. Experiments revealed a waveform curves of antibody levels to different porin forms reflecting the successions of immunoglobulin classes and superposition of independent responses to various antigen determinants. The dynamic of immune response to porin depends on the molecular form and dose of the antigen. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 10, pp. 437–440, October, 1999     

Termination of Immunologic Tolerance with Tolerated Antigen Complexed with Specific Antibody

Female NTH albino and BALB/c mice were tolerized with chicken egg lysozyme or capsular polysaccharide of Diplococcus pneumoniae type III (SIII). Isolated spleen cells from tolerant mice were incubated in vitro with antigen-antibody complexes formed with the tolerated antigen and specific xenogeneic, allogeneic and syngeneic immune serum. The spleen cells were washed and returned to the individual donor mice and their immune response to the tolerated antigen was assayed. In all cases one or more concentrations of specific antibody when complexed with the tolerated antigen was able to terminate the tolerant state. The successful use of specific antibodies raised in a syngeneic strain of mice (BALB/c) to terminate immunologic tolerance in other BALB/c mice with demonstrated T and B cell tolerance, precludes T cell recognition of a foreign antibody as a mechanism to explain these results.     

The immune reponse to type III pneumococcal polysaccharide in mice with malaria.

The immune response of BALB/c mice to type III pneumococcal polysaccharide (SIII), as measured by splenic PFC, was abolished at the height of an acute self-limiting attack of malaria caused by the murine plasmodium P. yoelii, over a wide range of antigen doses. The response to antigen, given at various times after clinical recovery, gradually reappeared, but did not reach normal levels until 12 weeks after the injection of the parasite. A second injection of P. yoelii given 1 hr before SIII caused a moderate degree of depression, although in this case the plasmodium does not multiply. In chronic malaria the response to SIII was also very poor. Short term under-nourishment was found to reduce only slightly the response to SIII.     

Rate of development of immunological memory and characteristics of the secondary immune response in mice of strains with high and low reactivity

Immunological memory for sheep's red blood cells develops in mice of strains CBA and DBA/2 and (CBA×C57BL/6)F₁ hybrids 24 h after injection of a small dose of the antigen, but 48 h after injection in C57BL/6 mice. The level of the secondary immune response in CBA, C57BL/6 and F₁ hybrids is significantly higher than in DBA/2 mice. Maximal production of antibody-forming cells in the spleen of the CBA mice is observed after two injections of small doses of the antigen. By contrast to this, to obtain a marked immune response in the case of adoptive transfer of spleen cells of C57BL/6 mice a second injection of a large dose of antigen is required.Laboratory of Immunogenetics, Institute of Medical Genetics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. H. Zhukov-Verezhnikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 10, pp. 454–457, October, 1977.     

Major histocompatibility complex regulation of the class of the immune response: the H-2d haplotype determines poor interferon-gamma response to several antigens

The lymph node cells of CBA (H-2k), but not BALB/c (H-2d) mice, release interferon (IFN)-gamma into the supernatant when immunized with picryl chloride epicutaneously and then exposed to antigen (haptenized cells) in vitro 4 days later. The failure in IFN-gamma production maps to the major histocompatibility complex (MHC; H-2d) in the congenic BALB/c, BALB/k and BALB/b mice. The evidence that this is an MHC regulation of the class of response to a range of antigens and not a classical Ir gene effect is (a) the difference is seen with several antigens including picryl chloride, "oxazolone" and purified protein derivative of tuberculin and (b) BALB/c mice, which fail to produce IFN-gamma, show excellent contact sensitivity to picryl chloride. It was also found that the crosses between responder and nonresponder strains (CBA x BALB/c)F1 respond to antigen on responder cell but not on nonresponder cells. This influence of MHC on the class of the immune response is a possible basis for some of the associations of MHC with disease.     

Immune response of mice of various inbred lines toClostridium oedematiens toxoid

Mice of various inbred lines were immunized intradermally withClostridium oedematiens toxoid. The immunization was repeated 30 days later. The titer of antibodies against toxoid was determined by the passive hemagglutination test in the blood of the mice 20 and 30 days after the first and 10 days after the second immunization. The maximal response to primary immunization was recorded in C3H mice, the minimal in DBA/2 mice, with a more than 30-fold difference. The remaining tested lines of mice (A, CBA, BALB/c, AKR, C57BR) occupied an intermediate position. After the second immunization the differences were reduced. The existence of genetic control of the immune response to this particular antigen is postulated in mice.Translated from Byullenten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 2, pp. 198–200, February, 1977.     

Phytohemagglutinin stimulation of enhanced immunoglobulin G production in mice inoculated with type III pneumococcal polysaccharide.

In BALB/c mice previously inoculated intraperitoneally with an immunogenic dose of the T-independent antigen type III pneumococcal polysaccharide, the intravenous administration of the T-cell activating agent phytohemagglutinin P causes a pronounced increase in the number and relative proportion of immunoglobulin G-producing cells. These results, detected by a modified hemolytic plaque assay, were supported by finding increased levels of serum immunoglobulin G anti-type III pneumococcal polysaccharide in the treated mice. A comparable stimulation of immunoglobulin G antibody-producing cells was not induced in phytohemagglutinin P-treated nude mice, indicating that the change in class of the predominant antibody is attributable to the activation by the phytohemagglutinin P of a T-cell population. Under the conditions of these experiments, phytohemagglutinin P also promotes a progressive suppression of the antibody-forming cells during the response to type III pneumococcal polysaccharide.     

Antibody Response to Pneumococcal Polysaccharide Vaccine in Young, Adult and Old Mice

The anti-pneumococcal antibody response was studied in young (5-week-old) and adult (10-week-old) BALB/c and CBA/J mice and in adult (9–10-week-old) and old (12-, 18- and 24-month-old) AB6F₁ and B6D2F₁ mice after s.c. immunization with a 23-valent pneumococcal polysaccharide vaccine. Both young and adult mice showed a significant IgM antibody response to the vaccine 6 days after immunization with 111 /ig antigen. There were significant immune responses to serotypes 1, 2, 4 and 7F in contrast to small responses to serotypes 14, 19F and 23F after immunization with the vaccine. One month after immunization, there were only marginal differences in IgM anti-pneumococcal antibody levels to the vaccine (anti-PPS) between immunized and unimmunized BALB/c mice, whereas in CBA/J mice the anti-PPS remained higher in immunized than in unimmunized mice. Immunization of old mice induced a significant IgM antibody response 6 days after immunization, but the anti-PPS thereafter decreased rapidly towards preimmunization values in AB6F₁ mice. A significant IgG anti-PPS was not detected in any of the mice studied. The IgA anti-PPS tended to vary over time with no consistent pattern. It is important to carefully consider age and strain of the mice used when studying the immune response to pneumococcal polysaccharide antigens.     

Immunogenicity of gonococcal Gc2 polysaccharide: comparative studies with pneumococcal type III polysaccharide and Salmonella typhosa Vi antigen.

A plaque assay technique was used to assess the immunogenicity of a gonococcal cell wall polysaccharide (Gc2 antigen) in BALB/c mice. The Gc2 antigen was shown to be immunogenic, and the kinetics of the response differed from that of a pneumococcal polysaccharide (SSS-III) and a polysaccharide antigen of Salmonella typhosa (Vi antigen). In addition, using antithymocyte sera, the T-lymphocyte dependency of these antigens was investigated. The immune response to the Gc2 antigen was demonstrated to be dependent on a population of helper T cells, whereas the response to SSS-III appears to be regulated by suppressor T cells. There appears to be marked differences in the immune response of mice to different bacterial polysaccharides.     

The capsule of Bacillus anthracis behaves as a thymus-independent type 2 antigen

Bacillus anthracis elaborates a homopolymeric capsule composed of gamma-D-glutamic acid residues. Mice were immunized with formalin-fixed encapsulated B. anthracis bacilli, and the serum antibody response to a gamma-D-glutamyl capsular epitope was measured. Antiglutamyl antibodies were elicited in athymic BALB/c Nu/Nu, BALB/c Nu/+, and CBA/J mice but not in CBA/N xid mice. These response patterns define the capsule of B. anthracis as a thymus-independent type 2 antigen.     

Effect of cocaine on the immune response and host resistance in BALB/c mice

This study focuses on the effect of varying regimens of cocaine administration on three parameters of the immune response: antibody production, resistance to infection by Streptococcus pneumoniae following immunization, and resistance to tumors. The effect of cocaine on antibody production of female and male BALB/c mice was investigated to both a T-independent (pneumococcal polysaccharide type III [SSS-III]) and a T-dependent antigen (the 2,4-dinitrophenyl ligand [DNP]). It was found that high doses of cocaine injected 3 times/day prior to SSS-III resulted in a small rise in antibody levels in male mice. Low doses given for 4 days prior to or subsequent to SSS-III injection had no effect on the antibody response nor on the susceptibility to infection by live S. pneumoniae. High dosages of cocaine administered 3-5 times/day had no effect on the anti-DNP immune response of male mice but resulted in an almost 2-fold increase of anti-DNP plaque-forming cells in female mice.     

Age-related difference by IgG subclass in the response of mice to allogeneic spleen cells.

Inbred mice of various strains, 3–4 months of age and 16 months or older, were given primary injections of allogeneic spleen cells to observe the time of appearance and relative levels of alloantibodies of IgG1 and IgG2 class. In 3–4 month old C3H mice injected with BALB/c spleen cells, IgG2 alloantibodies were present on day 6, before the appearance of alloantibodies of IgG1 class. The IgG2 class alloantibodies then continued to increase in level until day 12. IgG1 class antibodies, which appeared after day 6, also increased during this period. When C3H mice at 16 months of age were similarly injected a difference in response was found in that IgG2 class alloantibodies did not increase in level after the appearance of those of the IgG1 class, but IgG1 class alloantibodies increased until day 12, as in the young mice. At intermediate ages smaller effects in the same directions were observed. Young BALB/c mice injected with C3H spleen cells gave responses similar to those of the young C3H mice. At 16 months, however, the response was not different from that of the young BALB/c mice. At older ages (19–25 months) the response of BALB/c mice was similar to that of the 16-month-old C3H mice described above. CBA mice of various ages which were injected with BALB/c spleen cells showed effects similar to that of the BALB/c mice in that a change of response seen in the older CBA mice, similar to that of the other two strains, did not appear at 16 months of age but did appear later (20–24 months).     

Immunopathological aspects of Plasmodium berghei infection in five strains of mice. I. Immune complexes and other serological features during the infection.

The development of circulating immune complexes was studied in mice of the BALB/c, A/J, OF1, CBA and C57B1 strains infected with P. berghei. Complexes were evaluated in relation to levels of parasitaemia, soluble antigen, specific antibody and C3. Susceptibility to infection was greatest in BALB/c, A/J and OF/a mice. The maximum parasitaemia was 30% in CBA and 70% in all other strains. Levels of soluble antigen paralleled those of parasitaemia. Specific antibody was detected in all strains, but the titre continued to rise throughout the infection only in CBA mice. Circulating immune complexes occurred in mice of all strains from day 6; the level fell after day 9 in C57B1 whereas it was maintained in CBA mice. The development of immune complexes was associated with marked depression of C3 levels in all except CBA mice, in which a transient reduction was followed by recovery. Partial characterization of the complexes showed that IgM-containing complexes appeared earliest and reached highest levels in BALB/c mice while in CBA mice, IgM complexes were found in lesser amounts and the level fell in late infection. IgG complexes rose throughout infection in CBA and fell in later stages in BALB/c and C57B1 mice. In nude BALB/c mice, immune complexes were usually not detectable and only low levels of antibody of IgM class were produced. Differences in mortality pattern could not be related to any single serological factor.