短小棒状杆菌对小鼠实体瘤及免疫功能的影响

目的研究短小棒状杆菌对H22荷瘤小鼠及其免役功能的影响。方法采用抑瘤实验、小鼠碳廓清实验、MTT法观察短小棒状杆菌对小鼠实体瘤和免疫功能的影响。结果短小棒状杆菌可使荷瘤小鼠的碳廓清指数、胸腺指数、脾脏指数升高,并由刀豆蛋白(ConA)、脂多糖(LPS)诱导体外小鼠T、B淋巴细胞增殖,能够抑制小鼠实体瘤生长。结论短小棒状杆菌能够增强小鼠机体免疫功能,并有抑瘤作用。     

A phase I study of intrapleural Corynebacterium parvum after complete resection of bronchogenic carcinoma

Summary Corynebacterium parvum (C. parvum) was instilled into the pleural space via the chest tube in 11 patients after curative resection for lung cancer. Doses were escalated from 20–70 mg in approximately every third patient in an attempt to determine the maximum tolerated dose. Fever and chest pain were the only toxicities encountered; severity and duration were not dose-related. Six of seven surgical stage I patients were alive and free of recurrence with a median follow-up of 2 1/2 years. A single patient developed light-chain-producing multiple myeloma 1 year after C. parvum injection.American Cancer Society Clinical Fellow     

Splenic suppressor macrophages induced in mice by injection of Corynebacterium parvum.

Spleen cells from C57BL/6N mice injected with killed Corynebacterium parvum (CP) had a marked growth inhibitory effect on the in vitro proliferation of RBL-5 murine lymphoma cells. It was most marked 12 to 14 days after injection and was usually no longer detectable later than 21 days. It could be demonstrated at effector cell to target ratios between 20:1 and 5:1 at which normal spleen cells had a growth-promoting effect. Addition of CP to an in vitro mixture of spleen cells and tumor cells augmented the inhibitory effect of spleen cells from CP-injected mice although it conferred no inhibitory potential on normal spleen cells. Growth inhibiton by CP spleen cells was not mediated by T cells and various depletion experiments suggested that the effector cells of the phenomenon were macrophages. Spleen cells of CP-injected mice also showed strongly depressed responses to the T cell mitogens PHA and Con A and suppressed the mitogen responses of syngeneic normal spleen cells. The characteristics of the suppressor cells mediating this effect appeared to be very similar to those inhibiting lymphoma cell growth. The responses to LPS were also strongly suppressed in mice injected with 2.1 mg of CP. However, after injection of one-tenth of the dose a relative sparing of the LPS response was noted, whereas the PHA response was still suppressed.     

Increased susceptibility to Escherichia coli infection in mice pretreated with Corynebacterium parvum

The contribution of activated macrophages to protection against Escherichia coli was studied in mice treated intravenously with Corynebacterium parvum 7 days before infection. C. parvum-treated mice showed increased phagocytic activity and enhanced resistance to Listeria infection. In contrast, these mice showed increased susceptibility to a subsequent challenge with E. coli that correlated closely with a reduction in the LD50 of lipopolysaccharide (LPS) in these mice. The peritoneal macrophages obtained from C. parvum-treated mice had a strong ability to phagocytize and kill E. coli in in vitro experiments. A rapid decline in the number of bacteria in the liver of C. parvum-treated mice was observed in the early period of infection. However, the number of bacteria in liver and spleen increased progressively to a lethal dose from 6 hr after infection. At this time, a significant increase in beta-glucuronidase, a lysosomal acid hydrolase, was found in the serum of these mice. In vitro experiments revealed that the peritoneal macrophages from C. parvum-treated mice were highly susceptible to the cytotoxic effect of LPS after 6 hr of incubation with LPS. It is suggested that the hypersensitivity of activated macrophages to the cytotoxic effect of endotoxin derived from E. coli may be partly responsible for the increased susceptibility of C. parvum-treated mice to E. coli infection.     

Effects of Corynebacterium parvum on depressed immunological reactions in EL-4-tumor-bearing mice

Summary Effects of Corynebacterium parvum on the development of plaque-forming cells (PFC), cell-mediated cytotoxicity (CMC), and delayed footpad reaction (DFR) to chicken erythrocytes (CRBC) were investigated in EL-4-bearing syngeneic mice. PFC, CMC, and DFR responses after the primary immunization were suppressed in tumor-bearing mice and restored by C. parvum treatment. PFC and CMC responses in tumor-bearing mice were restored by the transfer of spleen cells of C. parvum-treated normal mice. Such powers of recovery were abrogated by the removal of glass-adherent cells but not by the removal of -positive or Ig-positive cells. DFR was suppressed not only in the primary but also in the secondary immunization, in contrast to PFC and CMC; the secondary responses of these types were not suppressed in tumor-bearing mice. Positive DFR was not elicited in tumor-bearing mice after adoptive transfer of sensitized lymphocytes from normal immune donors. The DFR became positive in such tumor-bearing recipients when they were treated by C. parvum. Macrophage functions in the induction phase of the immune response as accessory cells and in the expression of DFR as secondary cells appear to be suppressed in tumor-bearing mice and restored by C. parvum.     

Antitumor activity of Corynebacterium parvum administered into the pleural cavity of mice

Summary We investigated the efficacy of intrapleurally (IPl) injected 0.25 mg Corynebacterium parvum (CP) against pulmonary tumor deposits of four syngeneic murine tumors, C3Hf/Bu or CBA fibrosarcoma (FSa) or mammary carcinoma (MCa), and compared its efficacy with that of intravenous (IV) CP. A mode of action of CP given by IPl administration was also studied. When given to mice prior to the IV inoculation of tumor cells, IPl and IV CP reduced the number of metastases of C3Hf/Bu-FSa and CBA-MCa equally well. Intrapleural injection of CP within a week after tumor cell inoculation was more effective against metastases in the lung than IV CP. A combination of IPl and IV administration of CP was more effective in the therapy of lung metastases of CBA-FSa and CBA-MCa than either single treatment alone. Intrapleural and IV injections of CP augmented concomitant immunity to artificially produced lung metastases of C3Hf/Bu-FSa. Intrapleural CP was less effective than IV CP in inducing complete regression of the SC growing C3Hf/Bu-FSa.In contrast to IV CP, IPl CP did not markedly influence the spleen weight and cellularity. It was also less effective in increasing the liver weight. However, CP by either route of injection led to a significant increase in the lung weight. CP injected IPl, but not IV, caused a significant increase in the number of nucleated cells in the pleural cavity of mice. More than 90% of these cells were macrophages, and they were found to be cytotoxic for in vitro cultures of CBA-MCa cells. Activated macrophages also mediated in vivo antitumor resistance, as shown by the abolition of this resistance by treatment of the mice with carrageenan.     

Prostaglandin synthesis in spleen cell cultures of mice injected with Corynebacterium parvum.

Spleen cells of C57BL/6 mice produced high amounts of PGE in vitro when tested 5 to 10 days after injection of heat-killed C. parvum organisms. Little or no PGE was produced by spleen cells from untreated mice or from mice injected with a strain of coryneform bacteria that does not stimulate the lymphoreticular system of mice. Significant release of PGE from spleen cells of C. parvum injected mice could be detected as early as 30 min after initiating the cultures and maximal levels were usually seen after 48 hr. Treatment by indomethacin completely abolished this PGE production. Removal of the adherent population from the spleen cell suspension resulted in markedly decreased levels of PGE, but PGE release of the remaining population was never completely abolished. These data suggest that the cells responsible for most of the PGE synthesis in this system were adherent cells, presumably macrophages. The levels of PGE produced in spleen cells of C. parvum-treated mice were further increased by in vitro addition of C. parvum. This effect could also be observed after addition of zymosan particles indicating that it was not an immunologically specific effect. The reported data suggest that prostaglandins may represent important mediator molecules of the described immunostimulatory and immunosuppressive effects of C. parvum.     

Immunomodulation by Corynebacterium parvum in normal humans

Four normal human donors were immunized with 2 mg of heat-killed Corynebacterium parvum by subcutaneous and intracutaneous injections, and lymphocyte surface markers, antibody-dependent (ADCC), and spontaneous cell-mediated (SCMC) cytotoxicities followed for a 28-day period. Although no changes were observed in the relative proportions of B, T, and Fc receptor-carrying lymphocytes, two T cell subpopulations, namely, the autologous rosette-forming cells and active rosette-forming cells, both exhibited significant increases. Significant increases were also demonstrated in the proportion of monocytes carrying Fc receptors and in the proportion of monocytes phagocytizing Latex beads. Although no consistent changes could be found in ADCC against the P 815 mastocytoma cell line, the SCMC against both the myeloid leukaemia line K 562 and the lymphoma line RAJI was found to be elevated as early as 6 hr post-vaccination. The possibility that the enhanced SCMC activity was induced in vivo by interferon was supported thus: 1) Enhancement of SCMC in vitro by interferon was abrogated by the vaccination. 2) Serum interferon determinations showed significant increases in parallel with the lack of in vitro SCMC enhancement.     

Splenocytes from tumor-bearing Corynebacterium parvum treated mice maintain interleukin-2 and -3 activity

Summary Systemic and local administration of the bacterium Corynebacterium parvum (more accurately known as Propionibacterium acnes) is reported to exert antitumor action via activated macrophages or short-lived cytotoxic T lymphocytes (CTL), respectively. This study examined the effect of C. parvum treatment on resulting in vitro interleukin levels, which are components in the sequence of events leading to the development of effective CTL. C. parvum administration prevented palpable fibrosarcoma development. This was concomitant with restoration and maintenance of normal interleukin-2 (IL-2) and interleukin-3 (IL-3) levels and prevention of suppressor cell development in mice injected with both tumor cells and vaccine. Our finding of C. parvum-induced maintenance of IL-2 and IL-3 levels and apparent lack of suppressor cell formation lends support to the idea of local C. parvum antitumor action possibly being mediated by CTL arising via the interleukin cascade.     

The antineoplastic effect of Corynebacterium parvum modified by lysozyme treatment

Summary The study was conducted to evaluate the possibility that host factors may modulate the antitumor effect of C. parvum (CP). Heat-killed CP after being subjected to limited digestion by lysozyme (L-CP) produced a superior effect on the inhibition of local growth of murine mammary carcinoma (CD ₈ F ₁ ) comparsed to unmodified CP. The median survival time (MST) of tumor-bearing animals that were treated with L-CP was slightly but not significantly increased over that of the immunotherapeutic response to unmodified CP. This modified CP was capable of reducing the incidence of early death in tumor-bearing animals. The modified CP was also more toxic than the unmodified. In contrast, mixed glycosidase had no effect on the antitumor potency of CP. The usefulness of enzymatic modification of whole CP in improving the tumor-therapeutic potency of CP is discussed.     

Immunotherapy of lung cancer with Corynebacterium parvum

Summary Thirty-one patients with inoperable carcinoma of the lung, excluding oat-cell carcinoma, were randomized to receive either chemotherapy alone, with methyl CCNU and vinblastine every 6–8 weeks (15 Pts) or such chemotherapy plus immunotherapy with IV infusions of Corynebacterium parvum (16 Pts). Prior duration of the disease was longer, and more patients had received previous therapy, in the immunotherapy group; these groups were otherwise very similar. In vitro lymphocyte response to phytohemagglutinin did not change significantly in either group, but the weaker response to Varidase declined in both groups after chemotherapy. An increased baseline level of circulating B lymphocytes was sharply reduced in the C. parvum group. There were no differences in -globulins or delayed skin test responses between immunotherapy and control patients at entry into this study or on follow-up. Median survival from entry was longer in the immunotherapy group (6 months) than in the control group (3 months), but this difference was not statistically significant and only two patients in each group lived for more than 11 months. It is conceivable that more benefit from C. parvum might have been recorded had more effective chemotherapy been available.     

Inhibition of cell-mediated cytotoxicity by Corynebacterium parvum

Summary We investigated the effect of altering dose and route of Corynebacterium parvum (C. parvum) administration on the adjuvant's inhibition of cell-mediated cytotoxicity (CMC). Primary in vivo and secondary in vitro CMC of C57B1/6 mice alloimmunized to P815 were depressed if C. parvum was administered systemically (IV or IP) but not when it was given SC. Similarly, only systemic C. parvum generated cells capable of suppressing in vitro CMC. Primary and secondary CMC in spleen was equally inhibited by 700 and 70 g, whereas suppressor cell activity was marked with 700 g and minimal with 70 g. Administration of C. parvum SC admixed with alloantigen resulted in early enhancement and late depression of primary CMC. Secondary CMC was depressed but suppressor activity was absent. Dissociation of CMC depression from suppressor cell generation indicates that these phenomena can be separated under certain conditions.     

Biological effects of Corynebacterium parvum. IV. Adjuvant and inhibitory activities on B lymphocytes

The PFC response to the thymus-independent antigen SIII (type 3 pneumococcal polysaccharide) was amplified in mice injected 4 days previously with killed Corynebacterium parvum. This adjuvant activity was demonstrable with high (2–50 μg) but not low (0.1–0.5 μg) doses of SIII. Induction of tolerance was unaffected. Depression of the response resulted from simultaneous injection of SIII with either C. parvum or Bordetella pertussis, while prior treatment with the latter was without effect. Responsiveness to SIII was transiently but potently suppressed in spleen cells transferred into lethally irradiated, C. parvum pretreated mice.Although C. parvum is an effective B cell adjuvant, other data imply that it acts indirectly on these lymphocytes. It is argued that both adjuvant and suppressive activities of C. parvum on the B cell response to SIII are most probably mediated by activated macrophages.     

Active Immunotherapy with Corynebacterium parvum and Chemotherapy in Murine Fibrosarcomas

Corynebacterium parvum used alone to enhance immunological reactivity produced transient inhibition of the growth of chemically induced isogenic mouse tumours. Attempts were made to combine C. Parvum with cyclophosphamide to see whether this would increase the latter''s effectiveness in inhibiting early but established tumours. Of the various regimens tested, the administration of the C. parvum 12 days after a single dose of chemotherapy produced dramatic inhibition of tumour growth and resulted in complete and lasting regressions in up to 70% of tumour-bearing animals. The most important variable in this regimen is the time between the chemotherapy and the subsequent immunotherapy.It is possible that non-specific active immunotherapy with agents such as C. parvum may be a valuable adjunct to the conventional cyto-reductive treatments of cancer, but the time of administration of such therapy is probably critical for each tumour and for each chemotherapeutic regimen.     

Biological effects of the adjuvant Corynebacterium parvum. II. Evidence for macrophage-T-cell interaction

The mechanism underlying the markedly reduced PHA responsiveness of spleen and peripheral blood lymphocytes from Corynebacterium parvum-treated mice is due to inhibition of the responsive T-lymphocytes by C. parvum-activated macrophages. Inhibition is a result of a qualitative rather than quantitative change in the macrophage population and GVH-activated macrophages behave similarly. Corynebacterium parvum-activated macrophages need to be viable and will inhibit normal lymphocytes. This inhibitory effect appears to be mediated through cell-cell contact.     

Properties of an Antigenic Polysaccharide from Corynebacterium parvum

Corynebacterium parvum strain 10390 is an antitumor agent and stimulant of the reticuloendothelial system and produces a soluble antigen towards the end of its growth cycle. This material, which is a cell wall component and can also be released from the organism by acid or alkaline hydrolysis, has been purified. It is an acidic polysaccharide of molecular weight 100,000 to 150,000 and contains galactose, glucose, fucose, N-acetylgalactosamine, N-acetylglucosamine, uronic acids, sialic acids, and a small proportion of amino acids. The antigen gives a precipitin reaction with antisera raised against the whole organism and also binds to animal cells. The antigenic determinants are extremely resistant to oxidation, reduction, and enzymatic and chemical hydrolysis, but the single cell-binding site is destroyed by alkali and also by Helix pomatia digestive juice, alginase, and neuraminidase without substantially affecting the molecular weight. This site is inaccessible until the molecule is released from the cell surface. The possibility that the soluble antigen is the biologically active fraction of C. parvum is discussed.     

Failure of Corynebacterium parvum to inhibit antipyrine metabolism in man

Summary In animals, Corynebacterium parvum lowers the rate of drug metabolism and enhances the pharmacologic effect of drugs requiring hepatic microsomal enzyme activity for elimination. A pilot study was conducted to assess this drug interaction in patients given clinical protocol doses of C. parvum. In individual patients, C. parvum did not reduce microsomal drug metabolism as measured by antipyrine half-life. Conversely, antipyrine elimination appeared to be enhanced in 10 of 14 patients. Results from this small heterogenous patient group are not definitive, and further studies are needed to determine the clinical significance of the effects of nonspecific immunotherapy on drug metabolism.