Effect of Corynebacterium parvum on the immune response in guinea pigs. II. Passive transfer of enhanced anamnestic response and delayed hypersensitivity observed after treatment with Corynebacterium parvum]

After intradermal immunization with a mixture of Corynebacterium parvum (C. parvum) and ovalbumin guinea pigs show a markedly increased anamnestic response to an intradermal booster of ovalbumin as compared to controls treated with ovalbumin only. At the same time a reaction of delayed type hypersensitivity is observed in the treated animals, but not in controls. The enhanced anamnestic response as well as the posivitive skin reaction were transferred to strain 2 histocompatible guinea pigs by peripheral blood leukocytes as well as by peritoneal exudate cells. Passive transfer was not obtained after prior irradiation of donor animals.     

Prostaglandin-induced enhancement of the in vitro anamnestic response.

The ability of various prostaglandins (PGs) to affect the in vitro anamnestic immune response of keyhole limpet hemocyanin (KLH)-primed rabbit popliteal lymph node cells was investigated. Of the four PGs studied (PGA1, PGE2 and PGF2alpha), PGE1 was found to have a stimulatory effect, whereas PGA1, PGE2 and pgf2alpha were ineffective in stimulating or inhibiting the in vitro anamnestic response. Under the conditions studied, a 3.5-fold increase in antibody production was obtained in PGE1-treated, KLH-stimulated cultures. Maximum enhancement was obtained when 0.2 mug of PGE1 were added at the time of culture initiation and were allowed to remain in contact with the lymph node cells for 24 hours.     

Analysis of immune response of guinea pigs infected with Brucella melitensis

The dynamics of the first antigen specific stage of immune response to Brucella infection was experimentally studied with the method of binding adsorbed antigenic immunoreagents with lymphocytes. The study revealed that the content of antigen-binding lymphocytes (ABL) reached its maximum as early as on day 7 after infection, gradually decreasing afterwards (but even on day 90 ABL could be detected in the blood). The specificity of ABL was proved by the fact that they were absent in noninfected animals, while in the animals infected with Brucella their content was higher than that of ABL specific to Yersinia enterocolitica O9; Brucella-specific ABL bound Brucella lipopolysaccharide (LPS) more intensively than Yersinia LPS. The detection of Brucella-specific ABL was inhibited by Brucella LPS more actively than by Yersinia LPS. The evaluation of the affinity of ABL to homologous LPS, made by the ratio of binding immunoreagents of the same specificity, but with suboptimal and optimal specificity, proved that an increase in the avidity of ABL occurred in the dynamics of the infectious process, which corresponded to the increase of their specificity.     

Production of an antiviral factor by murine spleen cells after treatment with Corynebacterium parvum.

We have previously shown that injection of Corynebacterium parvum (CP) in mice protected them against lethal encephalitis induced by herpes simplex virus, (HSV). It is shown here that spleen cells of CP-injected mice in vitro produce a factor capable of inhibiting the replication of HSV in mouse embryo fibroblasts (MEF). A similar activity was produced after in vitro exposure of spleen cells from untreated mice to CP. CP was only slightly mitogenic in contrast to phytohemagglutinin (PHA), concanavalin A (Con A), and bacterial lipopolysaccharide (LPS) which were strongly mitogenic but did not induce antiviral activity high enough to be detected in the HSV-MEF system. The activity produced by CP-treated spleen cells appeared to be interferon since it was trypsin sensitive and species specific and not virus specific, and since preincubation of the cells was required to demonstrate an antiviral effect. However, the identity of CP-induced interferon with any of the previously described subclasses of interferon remains to be determined.     

Modification of the immune reaction by antigen-immunosuppressive-agent conjugates. IV. Studies on the specific suppression of humoral immune response in guinea pigs by antigen-immunosuppressive-agent conjugates]

Bovine gamma globulin (BGG) antigens were modified by the binding of 6-mercaptopurine and toluyl residues, and their influence on the humoral immune response in guinea pigs was investigated. The antigen-immunosuppressive agent-conjugates (AIC) were different, depending on the method used for their preparation and the number of coupled residues per one molecule of BGG. Conjugates denoted as MPI-n-BGG were prepared by special chemical binding of corresponding thioisocyanates. MPII-n-BGG were synthetized by acetylation, and MPIII-n-BGG conjugates, by reductive alkylation. Pretreatment of guinea pigs with MPIII-19-BGG, MPII-16-BGG resulted in a stimulatory effect on the subsequent humoral immune response induced by BGG application. A significant suppressive influence was detectable if the animals had been pretreated with MPII-6-BGG and MPI-26-BGG. MPI-13-BGG and MPI-36-BGG had no effect on the later induced anti-BGG antibody formation. The immune response against a second antigen (human serum albumin) was not influenced by this kind of pretreatment of the animals. Therefore it seems justified to conclude that both stimulatory and suppressive effects seen here were antigen specific and that both the method for chemical modification and the number of coupled 6-MP residues are very important for their effectivity.     

Effect of corticosteroids on human in vitro anamnestic response.

When human peripheral blood lymphocytes were cultured in the presence of the antigens streptodornase (SKSD) and allogeneic lymphocytes (MLC), increased rates of DNA synthesis were observed 4 to 6 days laer. If these cell cultures were continued to 12 days, the rates of DNA synthesis returned to control levels. On rechallenging these cells with the same antigens, an accelerated response was observed. The corticosteroid preparations hydrocortisone and methylprednisolone suppressed both of these responses and also impaired the ability of the 12-day cultures to generate cells responsible for the accelerated secondary response. Although cells cultured for 12 days in the presence of SKSD and hydrocortisone responded poorly to SKSD rechallenge, they responded well in MLC. Hence, the suppressive effect was not completely unselective.     

Delayed-type hypersensitivity and in vitro lymphocyte response in guinea pigs immunized with a live varicella vaccine

The relation of delayed type hypersensitivity (DTH) assessed by the Varicella-Zoster virus (VZV) skin test and lymphocyte transformation (LTF) with VZV antigen was investigated in guinea pigs immunized with live varicella vaccine virus, or heat-inactivated vaccine virus. Guinea pigs immunized with live varicella vaccine virus showed positive DTH and LTF responses to viral antigen as well as a neutralizing (NT) antibody response, while those immunized with heat-inactivated vaccine virus showed only an NT antibody response of the same degree as that to live vaccine virus. These results show the reliability of the skin test in assessing cell-mediated immunity (CMI) to VZV and the advantage of the live varicella vaccine over the inactivated one in immunizing guinea pigs.     

Humoral response in Treponema pallidum-infected guinea pigs. II. Circulating immune complexes and autoimmune responses

Guinea pigs of inbred strain 2 and of a strain deficient in complement component 4 (C4D) responded to intradermal infection with Treponema pallidum by production of antibodies to treponemal antigens, normal rabbit serum proteins, fibronectin, and creatine kinase and with formation of circulating immune complexes (IC). IC started to appear at low concentrations 1 mo after infection and increased between 3 and 5 mo post-infection. Antibodies to fibronectin appeared after the second month but were not detectable 30 days later. Antibody activity to creatine kinase was detectable at the fourth month and became significantly higher at 5 mo post-infection. Reinoculation with a dose similar to that used for primary infection caused a significant increase in all antibodies and IC. Dissociation products of IC formed after primary infection consisted predominantly of treponemal antibodies and antigens, whereas IC detected after reinfection consisted predominantly of antibodies and normal rabbit serum proteins. Antibodies to fibronectin and creatine kinase are considered autoantibodies, and the underlying mechanism responsible for their production in syphilis is discussed.