Effects of intravenous injection of two different strains of Corynebacterium parvum in the mouse.

A strain of C. parvum, CN6134, known to have antitumour activity, caused thrombosis in the sites where the organism is phagocytosed. It bound to macrophages in vitro and activated the alternate pathway of complement. A strain of C. parvum, CN5888, which fails to show antitumour activity, did not show thrombosis. It did not bind to macrophages or activate guinea-pig complement. It did, however, cause marrow infarction which seems to result from a diminished clearance of the organism from the circulation.     

Proliferative glomerulonephritis in mice given intravenous Corynebacterium parvum

Mice given a killed suspension of Corynebacterium parvum (C.p.) developed nephritis as part of an immune complex disease. The nephritis was dose-related. After a single dose of 70 microgram (a human-equivalent dose) or of 466 microgram there was a mesangiopathic glomerulonephritis and after repeated human-equivalent doses there was a mesangiocapillary glomerulonephritis. Antibodies to C.p. increased and circulating immune complexes were detected. Mice receiving repeated doses also developed an arteritis. Study of this model may help in the understanding of human immune complex disease and the pathogenesis of glomerulonephritis.     

The complications of intravenous Corynebacterium parvum infusion.

100 intravenous infusions of Corynebacterium parvum were given to thirty-six patients at a dose of 5 mg/m2. Fever and rigors occurred in all patients but these acute side effects were acceptable to all but two patients. Seventeen patients suffered a delayed fall in blood pressure, which was marked in eight of them but was generally well tolerated. One patient died 18 hr after infusion from extensive myocardial infarction. Herpes labialis complicated the first infusion in nine instances, which may reflect transient immunosuppression following immunotherapy.     

Mitogenicity of Corynebacterium parvum for mouse lymphocytes.

Corynebacterium parvum, a powerful in vivo immunostimulant, is shown to stimulate lymphocyte proliferation in vitro. C. parvum is mitogenic for B lymphocytes and does not stimulate T cells. The mitogenicity is dependent on the presence of macrophages. C. diphtheriae and another strain of C. parvum, both of which are devoid of adjuvant, RES stimulation, and tumour-inhibiting activities in vivo are nevertheless mitogenic. A lipid-free fraction from C. parvum, which retains in vivo immunostimulatory properties, is not mitogenic. Thus the mitogenic property is not correlated with the in vivo properties of C. parvum.     

Protection of mice against mouse hepatitis virus by Corynebacterium parvum.

C57BL/6 mice that are highly susceptible to infection with mouse hepatitis virus type 3 were protected against intraperitoneal viral infection by simultaneous intraperitoneal injection of Corynebacterium parvum. No protection was observed when C. parvum was given intravenously or when it was injected intraperitoneally 3 days before viral infection. Protective effects were, however, consistently found when C. parvum was given 2 h before or 2 h after viral infection. Activity was seen only against 10 50% lethal doses and not against 100 50% lethal doses. C. parvum also caused a significant decrease of virus type 3. These data suggest a direct effect of C. parvum on virus-susceptible cells. Injection of C. parvum in mice caused activation of natural killer (NK) cells and of interferon production. However, these two effects were equally demonstrable at high and low doses of C. parvum, whereas protection against mouse hepatitis virus type 3 was not demonstrable at low doses of C. parvum. Thus, antiviral protection may be dissociated from activation of NK cells and induction of interferon.     

Cytokine expression in the mouse brain in response to immune activation by Corynebacterium parvum

Cytokine expression in the brain has been suggested to mediate various sickness behaviors. Here we report that intraperitoneal injection of a Corynebacterium parvum antigen in C57BL/6 mice was followed by prolonged upregulation of cytokines in the cerebral cortex and subcortical structures in a time course that coincided with reduced spontaneous running activity.     

Effect of Corynebacterium parvum on the class and subclass of antibody produced in the response of different strains of mice to sheep erythrocytes.

Several strains of mice were injected with sheep erythrocytes (SRBC) using C. parvum as adjuvant. The adjuvant effects on the amounts of class and subclass of antibody produced were ranked in the order IgG2b greater than IgG2a and IgM greater than IgG1. In addition, these effects were shown to vary depending on the time of administration of C. parvum relative to antigen. C parvum was shown to have no adjuvant effect on the response of congenitally athymic mice when given at the same time as the antigen, SRBC. On the basis of the reported observations it is suggested that certain of the adjuvant effects of C. parvum require T-cell function.     

The characteristics of binding of Corynebacterium parvum to glass-adherent mouse peritoneal exudate cells.

Corynebacterium parvum, strain 10390, whole organisms were shown to bind to the surface of glass-adherent mouse peritoneal exudate cells in vitro. An HCl extract and a lipid extract of the organism were both capable of inhibiting this binding. The attachment of organisms was not affected by trypsin treatment of the cells, indicating that the plasma membrane receptor is not cell-bound antibody in nature. The binding was inhibited by various sugars, most of which are major components of the cell wall of C. parvum. Removal of divalent cations prevented binding. At room temperature some binding occurred in the presence of magnesium ions alone, whereas both calcium and magnesium ions were required at 4 degrees C. The possibility is discussed that the attachment of C. parvum to the plasma membrane of macrophages may lead directly to their activation.     

Characterization of mouse peritoneal exudate and associated leukocyte adherence inhibitory activity after intraperitoneal injection of either Bordetella pertussis or Corynebacterium parvum vaccines.

Bordetella pertussis and Corynebacterium parvum are commonly used immunopotentiating agents. To explore the inflammatory environment induced by these agents, the peritoneal exudate response in mice following intraperitoneal injection of B. pertussis (PV) and C. parvum (CV) vaccines was investigated. The PV-induced exudate isolated by lavage was characterized by an early neutrophil influx followed by enhanced accumulation of mononuclear cells and fluid protein. The CV exudate was principally mononuclear in nature and displayed fewer numbers of cells and less fluid protein. Both vaccines also enhanced the leukocyte adherence inhibitory activity (LAIA) of peritoneal fluid as measured in vitro. The development of exudate LAIA was T lymphocyte independent. A similar LAIA was demonstrated in nonimmune mouse plasma and serum. Exudate fluid and serum LAIA were heat stable and trypsin sensitive. These studies suggest that significant differences exist in the composition of the local tissue environment following PV and CV injection and that exudate LAIA is serum derived. Further studies in this direction should result in a better understanding of the ways in which inflammatory cells and fluid substances affect lymphocyte-macrophage interaction subsequent to adjuvant administration.     

Immune reactions in different mouse strains

The responses of immunocompetent cells to thymus-dependent antigen differ in mice of different strains. Immunization stimulated phagocytic activity of peritoneal macrophages in CBA/CaLac, DBA/2, and BALB/c mice and suppressed it in CC57W mice. By the formation of antibody-producing cells in the spleen in response to thymus-dependent antigen DBA/2 and CBA/CaLac mice can be classified as high responders, BALB/c mice as medium-responders, and C57Bl/6 and CC57W mice as low responders.     

The effect of intravenous Corynebacterium parvum on gut associated mononuclear phagocytes in normal and tumour bearing rats.

Corynebacterium Parvum, which has been used in the treatment of human colorectal cancer, probably exerts its action through cells of the mononuclear phagocyte system (MPS). In this study the effect of systemically administered C. parvum has been measured on gut associated MPS cells in normal and colorectal cancer bearing rats. MPS cells are not normally found in samples of lymph obtained after cannulation of the thoracic duct (TDC). However, after total extirpation of the mesenteric lymph nodes, TDC yields samples in which up to 5% of the total cell population appear to be MPS cells. This procedure has been carried out in adult Wistar rats enabling an in vivo study to be made on the effect of C. parvum treatment on the effluent gut cells. Measurements have been made both of the number of cells found in thoracic duct lymph and of their capacity to phagocytose sensitized sheep red blood cells. These measurements were repeated in a mesenteric lymphadenectomized group of rats which had also undergone induction of colonic cancer using dimethylhydrazine. C. parvum treatment did not effect total cell, or phagocyte numbers in thoracic duct lymph (TDL). However rats with colonic cancers showed a marked reduction in the numbers of phagocytic cells in TDL irrespective of C. parvum treatment.     

The effect of Corynebacterium parvum in influenza infected mice

This paper is the continuation of earlier studies on the effect of the killed suspension of Corynebacterium parvum in influenza virus infected mice. Our investigation showed the normalized effect of these drugs on disturbed function of cell mediated immunity during experimental influenza infection especially in phagocytic and bactericidal activity of granulocytes. The present experiments concern the explanation of these infection mechanisms. Intraperitoneal injection of Corynebacterium parvum stimulated spleen index. Foot pad test is higher than in comparatively treated BCG group. The pathomorphological analysis of the spleen, thymus and peritoneal lymph nodes points out to the multiplication of multiple lymph nodes sinus cells. Generally, C. parvum possessed protective effect in experimental influenza infection. We tested the following parameters: phagocytic and bactericidal activity of granulocytes, liberation of leukocytes migration inhibition factor (LIF).     

Pathogenicity of two Toxoplasma gondii strains in chickens of different ages infected via intraperitoneal injection

This experiment was conducted to investigate the pathogenicity of Toxoplasma gondii in broilers of different ages. Chickens at the ages of 7, 14, 21 and 28 days were injected intraperitoneally with 1 × 10⁸ tachyzoites of RH and JS strains of T. gondii, respectively. The clinical signs and death of chickens were recorded daily post inoculation. Serum samples were collected at days 0, 4, 11, 18, 25, 32, 39, 46 and 53 post infection to screen T. gondii circulating antigens (TCA) and T. gondii circulating antibodies (TCAb). The results showed that T. gondii infection of 7-day-old chickens caused death, even though the mortality rate of the JS strain (100%) was significantly higher than that of the RH strain (70%). Chickens at 14 days old showed only mild clinical signs, but no death. Neither clinical signs nor death were recorded in 21-day-old and 28-day-old chickens. TCA and TCAb became positive at days 4 and 11, respectively. Both the TCA and the TCAb of groups 21 days old (RH strain) and 28 days old (both RH and JS strains) decreased to a negative level earlier than the other experimental groups. Specific T. gondii DNA was detected by polymerase chain reaction in chickens that survived in the 7-day-old group (RH strain) and in all infected chickens of groups 14 days old and 21 days old injected with both strains. In the groups injected at 28 days old, three samples (RH strain) and one sample (JS strain) were found negative. The results indicated that the age of the chicken was an important factor affecting the pathogenicity of T. gondii and that these two strains of T. gondii displayed different virulence for chickens.     

Immunomodulation by corynebacterium parvum in two strains of guinea-pigs and the effect of cyclophosphamide.

An early immunosuppressive phase in the overall stimulatory effect of Corynebacterium parvum on the immune response of two strains of guinea-pigs to ovalbumin (Oa) is described. Boosters given soon after treatment with C. parvum elicited lower secondary responses than those given later on, the peak secondary antibody titre increasing with the interval between primary immunization and the booster injection. Transfer of blood leukocytes (lymphocytes) of immunized donors to virgin syngeneic recipients showed the cellular nature of this effect. Cells transferred from donors boosted on day 40 after primary immunization showed a mean adoptive response that was 3.5 times less than that of a similar number of cells from donors boosted on day 90. The increase in the immunocompetence and/or memory of the transferred cells was related to the moment of injection of the booster antigen (Ag) and not to the interval between priming and transfer, since cells transferred during the primary response failed to show a parallel increase. The early, lesser enhancement of the secondary response by C. parvum would thus appear to be due to a limitation of the number and/or immunocompetence of memory cells developing after the injection of Ag. A marked strain difference was observed in the response of strain 2 and Hartley guinea-pigs to immunization with C. parvum and Oa, delayed-type hypersensitivity (DTH) being relatively inhibited, and antibody (Ab) production relatively increased in the latter, the reverse being true of strain 2 guinea-pigs. The existence of a suppressor or regulatory mechanism sensitive to cyclophosphamide (Cy) during the development of DTH after C. parvum treatment was established. Cy pretreatment, 250-300 mg/kg i.p., of both strains led to the development of a larger number of positive skin tests in animals given C. parvum i.v. and Oa i.d. but not when C. parvum was injected i.d. mixed with Oa. The inhibition of the migration, in the presence of Ag, of peritoneal exudate cells of Cy-pretreated guinea-pigs was also accelerated and enhanced during the first week of the primary response after i.v. C. parvum.     

Modification of the antitumor action of Corynebacterium parvum by stress

Social grouping and isolation of mice, in the presence of an acute stressor, were found to differentially affect the antitumor action of the immunological adjuvant Corynebacterium parvum. Socially grouped DBA/2j mice were injected intradermally with P815 mastocytoma ascites cells. Half the mice had a threshold dose of C. parvum admixed with the P815 cells. Half the mice in each of those conditions were given acute, inescapable electric footshock. In a second experiment, the stressed mice were socially isolated prior to the acute stress. Tumor growth itself was not affected by the stress procedures. C. parvum inhibited tumor growth in non-stressed and socially isolated, stressed mice. However, social grouping selectively negated the C. parvum effect resulting in tumor growth and mortality equivalent to mice not given the adjuvant. Psychological factors may be important to the development of concomitant immunity and the efficacy of immunotherapies.     

Reaginic antibody production in different mouse strains

Mice of several strains were immunized with DNP-haemocyanin either with Al(OH)₃ or with Freund's complete adjuvant. Antibodies of immunoglobulin type γ₂ and γM were detected by passive haemolysis, γ_2a antibodies by PCA in guinea-pigs, γ₁ antibodies by PCA in mice using 2 hours of SP, and reaginic antibodies by PCA in mice using 72 hours of SP. Three of the six strains tested produced reaginic antibodies, i.e. SWR/J, C57BL/6J and A/HeJ, but only when Al(OH)₃ was used as adjuvant. All six strains produced γ₁ antibodies. Reaginic antibodies were separated by chromatography on DEAE-cellulose column.