Cell-mediated Immune Responses In Mice Research Articles

Recent studies on traditional Chinese medicinal plants

This review will discuss several new drugs which were discovered and developed in recent years from traditional Chinese medicines by the Shanghai Institute of Materia Medica. Huperzine A was isolated from Huperzia serrata, a plant used for the treatment of contusion, strain, haematuria, and swelling in Chinese folk medicine. Pharmacological studies have indicated that huperzine A has powerful and reversible anticholinesterase activity. Y-maze methods have shown that huperzine A improves learning and retrieval processes, and facilitates memory retention. Huperzine A is used to treat patients with myasthenia gravis and Alzheimer's disease in China. Sarmentosin, a cyanogenic glucoside was isolated from the whole plant of Sedum sarmentosium. This plant has long been used to treat hepatitis by folk medicine. Sarmentosin significantly lowers the SGPT level of patients suffering from chronic viral hepatitis, and shows a suppressive effect on cell-mediated immune responses in mice. The root of Aconitum is well known in traditional Chinese medicine. Many Aconitum alkaloids have been isolated. Most show potent bioactivities, but with severe toxicity. Recently, some alkaloids such as 3-acetylaconitine, lappaconitine, have shown significant anesthetic activity and exhibit a higher therapeutic index. Guan-fu base A was isolated from the tuber root of Aconitum coreanum. Guan-fu base A has antiarrhythmic action and is now in clinical trials. Drug Dev. Res. 39:147–157. © 1997 Wiley-Liss, Inc.

The antigen-specific cell-mediated immune response in mice is suppressed by infection with pathogenic lyssaviruses

Responsiveness of T cells (RTC) was studied in BALB/c mice intramuscularly infected with various lyssaviruses. After infection by this peripheral route, two types of viruses could be classified according to their effects: 1) pathogenic viruses, including fixed rabies Pasteur virus (serogenotype 1) and wild viruses belonging to serogenotype 1 (from a rabid fox in France and from a cow infected by a vampire bat in Brazil) or to serogenotype 5 (European bat lyssavirus 1); and 2) non-pathogenic viruses, including Mokola virus (serogenotype 3). RTC was tested by analysing in vitro the capacity of splenic T cells from infected BALB/c mice to produce cytokines after antigenic (purified lyssavirus antigens) or polyclonal stimulation (concanavalin A). Cytokine production was followed by assaying the biological activity of interleukin-2 and by testing for interleukin-2, interleukin-4 and interferon-gamma (IL2, IL4 and IFN gamma ) messenger RNAs (mRNA) by transcription into complementary DNA and amplification by the polymerase chain reaction. The initial biologically active IL2 and cytokine mRNA production was observed in mice infected with pathogenic or non-pathogenic lyssaviruses. Only mice with symptoms (infected with pathogenic viruses) lost the capacity to produce cytokines in vitro after antigen-specific stimulation. No such loss was observed after polyclonal stimulation. In mice peripherally infected with non-pathogenic viruses, no loss was observed after stimulation with lyssavirus antigens. Thus, infection with pathogenic lyssaviruses by the peripheral route induces in BALB/c mice a loss of T-cell responsiveness after antigen activation, but not after polyclonal activation.

Effect of mizoribine on effector T cell-mediated immune responses in mice.

T cells play a principal role in cellular immunity and govern the regulatory mechanism in humoral immunity. Therefore, T cells play a key role as either effectors or regulators in the immune network. Mizoribine (MZR), an immunosuppressive agent, suppresses both humoral and cellular immunity by acting on both T cells and B cells. In this study, we examined the effect of MZR on various effector T cell-mediated immune responses in mice. MZR prolonged skin graft survival and suppressed a localized graft-versus-host reaction (GvHR) and sheep red blood cell (SRBC)-induced delayed-type hypersensitivity (DTH) reaction. In a collagen-induced arthritic mice model, MZR reduced the arthritic index and the swelling of the hind limbs. Furthermore, MZR suppressed both bone damage and histopathological changes in the hind limbs. Interestingly, MZR markedly suppressed the DTH reaction to type II collagen (CII) but had no effect on anti-CII antibody levels in this arthritic model. In these models, effector T cells such as cytotoxic T lymphocyte (CTL) and TDTH cell play an important part in the development of these reactions. It is suggested that MZR inhibited these reactions via the inhibition of the effector T cell-mediated immune response. Therefore, it is also suggested that the suppressive effect of MZR on clinical rejection and autoimmune disease is based on its suppression of the effector T cell-mediated immune response, that is cellular immunity, in addition to humoral immunity.

Host range restricted, non-replicating vaccinia virus vectors as vaccine candidates.

Three model systems were used to demonstrate the immunogenicity of highly attenuated and replication-defective recombinant MVA. (1) Intramuscular inoculation of MVA-IN-Fha/np induced humoral and cell-mediated immune responses in mice and protectively immunized them against a lethal respiratory challenge with influenza virus. Intranasal vaccination was also protective, although higher doses were needed. (2) In rhesus macaques, an immunization scheme involving intramuscular injections of MVA-SIVenv/gag/pol greatly reduced the severity of disease caused by an SIV challenge. (3) In a murine cancer model, immunization with MVA-beta gal prevented the establishment of tumor metastases and even prolonged life in animals with established tumors. These results, together with previous data on the safety of MVA in humans, suggest the potential usefulness of recombinant MVA for prophylactic vaccination and therapeutic treatment of infectious diseases and cancer.

HIV gp120-Specific Cell-Mediated Immune Responses in Mice After Oral Immunization With Recombinant Salmonella

Salmonella is of great interest as a potential human immunodeficiency virus vaccine vector because of its ability to elicit potent mucosal and systemic immune responses when administered orally. To determine whether such a vaccine could elicit an immune response in mice, plasmids expressing HIV gp120-LAI were introduced into attenuated S. typhimurium. Three serial doses of 10(10) recombinant organisms were administered orally to BALB/c mice at 2-week intervals. Immunized mice but not control mice demonstrated proliferative T cell responses to gp120-LAI, comparable in magnitude to the proliferative responses to Salmonella antigens. Immunized mice had detectable serum and intestinal Salmonella-specific IgA and serum Salmonella-specific IgG. However, no gp120-specific antibody was detected in either serum or intestinal washes. These results indicate that live recombinant Salmonella-based vaccine constructs can induce HIV-specific cellular immune responses in vivo.

HIV gp120-Specific Cell-Mediated Immune Responses in Mice After Oral Immunization With Recombinant Salmonella

HIV gp120-Specific Cell-Mediated Immune Responses in Mice After Oral Immunization With Recombinant Salmonella

Influence of arecoline on immune system: III. Suppression of B cell-mediated immune response in mice after short-term exposure.

Arecoline, a major alkaloid of arecanut was examined to explore its modulatory influence on B cell-mediated immune response in a murine model system. The in vivo and in vitro effects were evaluated at sub-toxic concentrations of arecoline. The number of primary antibody forming cells (AFC) and hemagglutinating and hemolysis antibody titers to Sheep Red Blood Cells (SRBC) were evaluated in male mice. Arecoline exposure for a week invoked dose-dependent effect on primary antibody forming cells to SRBC with a maximum reduction at the dosage of 20 mg/kg bw, a moderate reduction at 10 mg/kg bw and no effect at 5 mg/kg bw dose level. HA and HL antibody titers to SRBC were suppressed markedly at arecoline dosage of 20 mg/kg bw and moderately at a dose of 10 mg/kg bw, given daily for 1, 2 or 3 weeks. The inhibitory effect of arecoline was not dependent on the duration of treatment. Like the primary antibody response, the secondary HA and HL antibody titers were also decreased after arecoline exposure. The administration of arecoline dosages 10 and 20 mg/kg bw daily for 4 days following SRBC immunization also, exerted dose-dependent suppression of primary antibody response. Similarly, when treated after 12 h following immunization, significant reduction in response was observed with arecoline dosage of 20 mg/kg bw. While moderate suppression of antibody response was noticed at the dose level of 10 mg/kg bw, there was no alteration in response at a dosage of 5 mg/kg bw.(ABSTRACT TRUNCATED AT 250 WORDS)

Suppression of the in vitro humoral immune response of mouse splenocytes by 7,12-dimethylbenz[ a]anthracene metabolites and inhibition of immunosuppression by α-naphthoflavone

Exposure to 7,12-dimethylbenz[ a]anthracene (DMBA) has been demonstrated by numerous investigators to result in suppression of both humoral and cell-mediated immune responses of mice and cultured splenocytes. The mechanism(s) of this DMBA-induced immunosuppression, however, is not well characterized. PAHs must be converted to reactive metabolites via cytochrome P450-dependent monooxygenase systems to exert their carcinogenic and mutagenic effects. Thus, we have hypothesized that immunosuppression seen upon exposure to DMBA may also be mediated by its reactive metabolites. The objective of this study was to determine if DMBA metabolites can suppress the in vitro, T-dependent humoral immune response to sheep red blood cells. Compounds were evaluated in the in vitro plaque-forming cell (PFC) response at concentrations of 10 −9 to 10 −5 m. DMBA and benzo[ a]pyrene (B[ a]P) were also evaluated for their ability to suppress the in vitro PFC response. Addition of either of these PAHs to splenocyte cultures produced a concentration-dependent suppression of the PFC response, in which B[ a]P was found to be 17.5-fold more potent that DMBA. These results are in contrast to those found in vivo, where DMBA has been shown to be more potent than B[ a]P at suppressing humoral immunity. The 3,4-diol metabolite of DMBA produced a concentration-dependent suppression (10 −8 to 10 −5 m) of the in vitro PFC response and was found to be 65-fold more potent than the parent compound DMBA. In contrast, the 5,6-diol metabolite of DMBA had no effect on the PFC response or cell viability. Both the 3-OH-DMBA and 7-hydroxymethyl-12-methyl-benz[ a]anthracene (7-OHMe-12-Me-BA) metabolites were found to be immunosuppressive at concentrations of 10 −6 m. Furthermore, suppression by 7-OHMe-12-Me-BA was observed at concentrations as low as 10 −8 m. Immunosuppression by the 7-Me-12-OHMe-BA and the di-OHMe-BA metabolites was only observed at high (10 −5 m) concentrations. The cytochrome P450 inhibitor, α-naphthoflavone (ANF), was utilized to determine if cytochrome P450-mediated metabolism is involved in DMBA-induced suppression of the in vitro PFC response. ANF (10 −5 m) reversed the DMBA-induced immunosuppression seen at 10 −5 m and attenuated the immunosuppression at 3 × 10 −5 and 10 −4 m. The results of these studies demonstrate that several metabolites of DMBA which can be generated by the cytochrome P450-dependent monooxygenase systems are immunosuppressive in the in vitro PFC response assay. Furthermore, the cytochrome P450 inhibitor, ANF, was able to reverse DMBA-induced immunosuppression. However, since the degree of DMBA-induced and B[ a]P-induced suppression of humoral immunity in vitro does not correlate with that observed in vivo, DMBA metabolites generated within the spleen may not be responsible for or may only partially contribute to the immunosuppression observed in the whole animal.

Humoral and cellular immune response in mice and dogs induced by a recombinant Echinococcus multilocularis antigen produced by a Salmonella typhimurium vaccine strain

A gene fragment (II/3-10) from the cestode Echinococcus multilocularis coding for a species-specific antigen was expressed in the live attenuated Salmonella typhimurium vaccine strain LT2 M1C. The recombinant vaccine (S. typhimurium + pVM II/3-10) was assessed for its potential to induce both a humoral and cell-mediated immune response in mice and dogs. Both subcutaneous and peroral administration of the vaccine resulted in antibody synthesis and lymphocyte priming of C57BL/6J mice against S. typhimurium antigens as well as against the recombinant E. multilocularis antigen. Two vaccinated (subcutaneous and peroral) dogs showed a strong humoral immune response to S. typhimurium antigens and to the recombinant E. multilocularis antigen, but proliferation of peripheral blood lymphocytes was not detectable against the bacterial S. typhimurium antigens. Regarding the recombinant E. multilocularis antigen, borderline lymphocyte proliferation was demonstrated following subcutaneous and none following peroral administration of the recombinant vaccine to dogs.

Protective immunity to experimental tuberculosis by mannophosphoinositides of mycobacteria.

Mannophosphoinositides isolated from mycobacterial cells were found to induce both humoral and cell-mediated immune responses in mice when injected as mannoside-methylated bovine serum albumin (MBSA) complexes. Immunization of mice with mannoside-MBSA complexes elicited significant protection against challenge with LD50 dose of M. tuberculosis H37Rv as revealed by high survival rate, low values of root-specific lung weight, lung densities and colony forming units recovered from lung, liver and spleen, compared to the nonimmunized group. These observations were further substantiated by histopathological studies. The protective immunity elicited by mannoside-MBSA complexes against challenge with M. tuberculosis H37Rv was mediated by the cooperation of T-B cells, as shown by the passive transfer of immune cells/sera into syngeneic sublethally irradiated recipient mice.

Serological, electrophoretic, and biological properties of Cryptococcus neoformans antigens.

We compared a cryptococcal culture filtrate antigen referred to as CneF with chemically defined cryptococcal antigen fractions isolated by Cherniak and co-workers by using double immunodiffusion gels, polyacrylamide gel electrophoresis, immunoblots, and footpad reactivity of immunized mice. The three previously described components of cryptococcal culture filtrates are a high-molecular-weight glucuronoxylo-mannan (GXM), which is the major constituent, a galactoxylomannan (GaIXM), and a mannoprotein (MP). In this study we demonstrated that CneF contained components which were serologically and electrophoretically similar to the three previously described cryptococcal culture filtrate fractions. The MP fraction elicited significantly stronger delayed-type hypersensitivity responses than did the GXM or GaIXM fraction when used in mice immunized either with the CneF in complete Freund adjuvant or whole heat-killed Cryptococcus neoformans yeast cells. These findings were confirmed when the footpads of immunized mice were challenged with GaIXM and MP preparations from a culture filtrate of a C. neoformans acapsular mutant that does not produce GXM. Thus, we concluded that the MP was the primary component recognized by the anticryptococcal cell-mediated immune response in mice.

The pathogenesis of interstitial pneumonitis induced by trehalose dimycolate. II. Reserpine prevents formation of lesions.

A single intraperitoneal injection of 10 micrograms of trehalose dimycolate (TDM) produced interstitial and hemorrhagic pneumonitis in C57BL/6 mice. As a part of an investigation of a possible role for cell-mediated immunity in the pathogenesis of this disorder, we found that reserpine, 3 mg/kg, given before, at the same time, or on Day 5 after administration of TDM, significantly reduced development of interstitial pneumonitis by Day 7. Smaller doses were less effective. Administration of reserpine, 3 or 2 mg/kg, 1 to 3 days after administration of TDM was lethal to most mice. Reserpine has been shown to inhibit expression of cell-mediated immune responses in mice, probably by causing intercellular release and degradation of vasoactive amines. Inhibition of pulmonary lesions by reserpine in TDM-treated mice suggests that a similar mechanism may be involved in the pathogenesis of TDM-induced lung injury.

Effector cell-mediated immune response in mice immunised with Salmonella.

Effector responses of cell-mediated immunity were studied in mice immediately and 3 months after immunisation with different kinds of salmonella vaccine. Whereas immunisation with live Salmonella enteritidis generated delayed hypersensitivity and activated macrophages, immunisation with formalised vaccine generated neither. Although activated macrophages had declined considerably by 3 months after immunisation, the delayed-hypersensitivity response had persisted. It was possible to generate activated macrophages in these mice by stimulation with sonicate antigens of the immunising strain or Salmonella paratyphic C. Delayed-hypersensitivity responses were elicited in mice, immunised with live S. enteritidis, by sonicate antigens of the homologous strains, S. gallinarum or S. paratyphi C.

Minimum peptide sequences necessary for priming and triggering of humoral and cell-mediated immune responses in mice: use of synthetic peptide antigens of defined structure.

Minimum peptide sequences necessary for priming and triggering of humoral and cell-mediated immune responses in mice: use of synthetic peptide antigens of defined structure.

Isolation of an enriched plasma membrane-subpellicular microtubule fraction of Leishmania mexicana amazonensis.

A cell fractionation procedure previously developed for Trypanosoma cruzi was applied to isolated the plasma membrane of promastigotes of Leishania mexicana amazonensis. The cell, swollen in an hypotonic mediun, were disrupted in the presence of a nonionic detergent and the membrane fraction isolated by differencial centrifugation. Electron microscopy showed that the fraction consisted of pieces of the plasma membrane associated with subpellicular microtubules. It was also shown that this fraction is able to induce cell-mediated immune response in mice.

Cell-mediated immune responses in mice infected with fonsecaea pedrosoi

Time course of cellular and humoral immune responses in mice infected with Fonsecaea pedrosoi was investigated by using an antigen prepared from culture filtrate of this fungus. Mice were infected by intravenous injection with yeast-like cells of the fungus. Viable fungus was recovered from the brain of the infected mice until the 36th day after inoculation, and from the other organs examined until 14th to 16th day. Inflammatory lesions were observed in the brain, lung, heart, liver, spleen, kidney and intestine during the first 30 days after inoculation. Macrophage migration inhibition factor response in these mice was insignificant until 8 days after inoculation. A significant response was developed at day 10 and persisted until day 63. This response returned negative by 95 days after inoculation. Lymphocyte transformation response of these mice was negative until 4 days after inoculation. At day 6 blastogenic index increased to 1.5, and at day 10, 14 and 16 the indices were 1.8, 2.4 and 1.7 respectively. Precipitin response to this fungus could not be detected in these mice until 16 days after inoculation, Positive results were obtained at day 21 and lasted until 51 days after inoculation. The precipitin titers, however, did not exceed one fold in any of these mice.

Cell-Mediated Immunity to Vesicular Stomatitis Virus Infections in Mice

The T cell-mediated immune responses of mice against vesicular stomatitis virus (VSV) were assessed by measuring direct primary foot pad swelling after local VSV infection and cytotoxic activity in spleens. The cytolytic activity was mediated by T cells since it was anti-theta + complement sensitive, was restricted by the K and D region but not the I region of H-2 and rapidly increased after 4 days but decreased 8 days after systemic or local infection. Cytolytic activity was virus-specific as reciprocally tested with VSV and vaccina virus immune T cells. Measurable activity on day 7 depended on infectious virus dose, virus virulence, and non-H-2 genetic background of the host. More than half of the cytolytic activity wasblocked specifically by either immune anti-H2 or rabbit anti-VSV antisera. Analysis of the kinetics of appearance of antigenic changes using metabolic inhibitors, revealed that the changes that rendered target cells susceptible to lysis after infection, occurred within the first hour after infection.

Examination of General and Tumor-Specific Cell-Mediated Immune Responses in Mice Bearing Progressively Growing Plasmacytomas 2

Examination of General and Tumor-Specific Cell-Mediated Immune Responses in Mice Bearing Progressively Growing Plasmacytomas 2

Inhibitory effect of alpha-globulins of different origin on the cell-mediated immune responses in mice.

alpha-Globulins prepared from different biological sources (human and bovine sera, mouse ascitic fluid, human ovarian cyst fluid) were separated by ion exchange chromatography on DEAE-cellulose into fractions A, B and C. Whereas fractions A and B had no immunosuppressive activity, fraction C injected into mice shortly before antigen administration, but not after, significantly inhibited the contact sensitivity to oxazolone. Single high doses were preferential over divided doses given on several occasions. In transfer experiments when cells of animals sensitized to oxazolone were injected into recipients treated previously with fraction C, the response was also inhibited. Fraction C, when administered into F1 hybrid mice before injection of parental splenocytes, prevented the development of GVH reaction. The authors point out that the critical concentration of alpha-globulins at the time of antigen administration is necessary to make T-lymphocytes hyporesponsive, and when once triggered by antigen they become refractory to alpha-globulins action.

Humoral antibody and cell-mediated cytotoxicity responses of mice to rat skin xenografts.

Aspects of both the humoral and cell-mediated immune responses of mice to rat skin xenografts were studied sequentially and quantitated. Antirat lymphocytotoxic and hemagglutinating antibodies were first detectable at 7 and 6 days, respectively, after primary grafting and their appearance in serum corresponded to the time of graft rejection. Lymphocytotoxic titers were low after primary grafting but increased greater than 4-fold after secondary grafting. Cytotoxicity of mouse spleen and axillary lymph node cells for 51Cr-labeled rat lymphocyte target cells was first detectable 5 days after primary grafting but was quite low showing a peak of only 7% specific 51Cr release 6 days after grafting. In contrast, the cytotoxicity of mouse spleen and lymph node cells for allogeneic target cells after primary skin allografting was significantly greater. It is suggested that the magnitude of the cell-mediated immune response to skin xenografts is less than the response to allografts and that the quicker and more vigorous rejection of skin xenografts is due to a larger participation of humoral antibody in the rejection process.