The Study Of Active Constituent Of Corynebacterium Parvum

Corynebacterium parvum(CP), which is one ofimmunomodulators has a variety of biological activities. It canregulate cellular immunity and humoral immunity by activatingmonocyte-macrophage, enhancing activities of NK cells,promoting the function of cytotoxic T lymphocyte, and inducing Tcell to produce IL-2 and the interferon. These functions arebeneficial to the effective elimination of viral antigens and variorusAg-Ab complex, and in this way the CP can increase the abilitiesresisiting diseases . Currently, it has been clinically used to treattumor, repeated upper respiratory tract infection, viral infection,feminine vulva floccosoids and psoriasis with the affirmableeffects.However, this product is whole cell preparation, and hasobvious side effects such as fever and injection area ache. which,as a matter of fact, limits its clinical application in a broader range.In this study, to ease the side effects, the active constituents of CPwere extracted by ultrasonic break-up, digestion and defating andits immunity activities were detected.1. The result of splenic activating test showed that thespleen-activated indexes of the purified CP group and the wholecell group increased markedly, in comparision with those ofnegative control group, reaching well over 1.5, and that the indexesrise with increase of doses. The purified group increased itsindex by approximately two times that of the whole cell group.This result indicates that the purified CP and the whole cellpreparations can obviously promote the macrophage proliferation.2. The result of the counts of splenocytes and the abdominalcavity macrophage suggested that both the purified CP and thewhole cell groups had a higher counts than that of the negativecontol group. Big cells (macrophage) were observed under anoptical microscope, and became adherent to the wall, spread andconfluent after five days of culture. This shows that the purified CPand whole cell preparations can markedly promote the macrophageproliferation.3. Through the experiment of the abdominal cavitymacrophage phagocyting chicken red blood cell, we observed thatthe phagocytosis percentage and indexes of the purified CP groupand the whole cell group were higher than that of the negativecontrol group. The purified CP can activate the macrophage andenhance its phagocytotic function.4. The result of NK cytotoxic activities determination showthat both the purified CP and the whole cell preparation have thesame function in enhancing the killing activties of NK cells. NKcell, being an important member of the non-special immune system,can kill the target cell directly without any MHC restriction.5. This result showed that the supernatant of splenocyte fromimmunized mice with the purified CP and the whole cell inducedL929 cells better growth in comparision with the negative control.This indicates that the purified CP and the whole cell preparationcan both induce the splenocytes to produce immunity-enhancedfactors. Through transformation of lymphocyte test, we observedthat supernatant of splenocyte of the purified group and the wholecell group can better promote the lymphocyte growth incomparison with the negative control group. It can be concludedthat the purified CP and the whole cell preparation may inducedthe production of such cytokine as IL-2 or IL-1. Because IL-2 isthe growth factor of lymphocytes, and IL-1 is able to coordinatewith lymphocytes activating.6. By stimulating the quadriceps of rabbits, we found thatpurified CP and the negative control group did not cause anyabnormal reactions such as hyperemia, necrosis, swelling, whereasin the whole cell preparation group there were obvious hyperemiaof a diameter of 2 cm, with central part necrosis. It is thereforeconcluded that the purified CP clearly decreases the stimulation tothe injected spots.7. The result of heat source test showed that the temperatureof the tested three rabbits in the purified group rose by less than0.6 ℃ after injection, while temperature of the other 3 rabbitsinjected with the whole cell preparation rose by 1.85 ℃ , 1.9℃ and0.85 ℃ respectively. This indicates that the purified CP does notcause any fever to the rabbits, whereas the whole cell preparationdoes lead to the heating reactions of the animals.8. Through the experiment of CP inhibiting tumor growth, weobserved that purified CP and the whole cell group both inhibitedtumor growth. The anti-tumor function may be carried out byactivating the macrophage, or by stimulating NK cell. Additionally,it may also function through cytokine (IL-2 and IFN). It was foundthat the activities of NK cell is closely related with the on-goinggrowth and metastasis of tumor and the survival of patients.The purified preparation of corynebacterium parvum made inthis study not only retains the effective activities of the whole cellpreparation, but also reduce obviously the side effects such as thepartial ache and fever. Moreover, in this research, we alsoobserved that the splenic and phagocytic index of the purifiedcorynebacterium parvum are clearly higher than those of the wholecell preparation. This indicates that the purified corynebacteriumparvum possesses a stronger phagocytotic function. The purifiedpreparation is easy and simple for production, inexpensive in cost,and has much less side effects as well. Therefore, it is accessibleto increase its clinical application in a wide range, and it will bringin great economic and social benefits. This purified preparation canbe developed into a novel product to replace the whole cellpreparation of corynebacterium parvum, and will surely bring ahealthy and happy life to the patients.