| Humen are being troubled by infectious diseases, and there are more limitation during the treatment courses for antibiotics to infectious diseases. Therefore, immunomodulation became hot point of prevention and therapy for infectious disease. There are many antigenic determinants in bacterial envolope,which are good immunomodulator. Some immunomodulators consisted of bacterial lysates were developed in overseas. We prepared bacterial lysates of Bifidobacterium bifidum, Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae on the basis of studies. And we detected the immunomodulatory effects in vivo and in vitro of individual bacterial lysates and mixed bacterial lysates.We got the Whole Peptidoglycan of Bifidobacteium bifidum(BbWPG) by chemo-enzymolysation. The bacterial lysates of Bifidobacterium bifidum(BbBL), Streptococcus pneumoniae(SpBL) and Haemophilus influenzae(HiBL) were prepared by sonicatolysation. The bacterial lysates of Staphylococcus aureus{SaBL) were extracted by enzymolysation. Lymphocyte transforming test (LTT) was used to detect the cellular immunomodulating effects of Bbwpc Bbbl, SaBL, BbWPG+SaBL,BbBL+SaBL, BbWPg+SaBL+SpBL+HiBL, BbBL+SaBL+SpBL+HiBL. The results showed that the concentration of BbWPG from 0.07g/L to 2.25g/L could not cause significantly cellular immunomodulating effects. The concentration of BbBL, 0.71g/L, could cause significantly cellular immunomodulating effects. Theconcentrating of SaBL, 0.012g/L, could cause significantly cellular immunomodulating effects, and when the concentration of SaBL was 0.195g/L it could cause the most significantly cellular immunomodulating effects. In the mixture of BbWPG+SaBL, the concentration of the group (40% BbWPG +60%SaBL)from 0.008g/L to 0.5g/L could cause significantly cellular immunomodulating effects and the concentration of other groups could cause significantly cellular immunoinhibiting effects. In the mixture of BbWPG+SaBL, the concentration of the group (40%BbWPG+60%SaBL) from 0.019g/L to 1.250g/L could cause significantly cellular immunomodulating effects, the concentration of the group(70%BbBL+30%SaBL) from 0.156g/L to 1.250g/L also could cause significantly cellular immunomodulating effects, and other groups of this mixture could not cause significantly immunomodulating effects. 95%(40/42) tested groups of the complex (BbBL+SaBL+SpBL+HiBL) could cause significantly cellular immunomodulating effects, and 36%(13/36) tested groups of the complex (BbWPG + SaBL + SpBL + Hibl) could cause significantly cellular immunomodulating effects. The Plaque Forming Cell assay was used to detect the ability of producing antibody forming cell. Both BbwpG(0.28g/L) and BbBL(0.70g/L) could produce more antibody forming cells(compared with the control group, p<0.01). BbWPG was better than BbBL. SaBi(0.1g/L) also could produce more antibody forming cells(compared with the control group, p<0.01). The mixture, BbBL+SaBL+SpBL+HiBL(1.45g/L, BbBL: SaBL: SpBL: HiBL =6. 25:6. 25:1:1, m/m), could produce more antibody forming cells(compared with the control group, p<0.05). The mixture, BbWPG+SaBL+SpBL+HiBL(1.45g/L, BbWPG: SaBL: Spbl: HiBL =8.33:12.5:2.33:1, m/m), could produce more antibody forming cells(compared with the control group, p<0.05). We observed the protectiveeffects of the mixture (BbBL+SaBL+SpBL+HisL, 1.45g/L, 0.2mL/mouse) and the mixture(BbWPG+SaBL+SpBL+HiBL, 1.45g/L, 0.2mL/mouse) on mice infected by Escherichia coli(E. coli) or Staphylococcus aureus(S. aureus). The survival rates of mice immuned with BbWPG+SaBL+SpBL+HiBL were 60%(infected by E. coli) and 91.7% (infected by S. aureus), which were compared with the control group,p<0.01 or p<0.05. The survival rates of mice immuned with BbBL+SaBL+SpBL+HiBL were 70% (infected by E. coli) and 72.7%(infected by S. aureus). No side effects on mice was observed after immuned withBbWPG+SaBL+SpBL+HiBL and BbBL+SaBL+SpBL+HiBL.After the protective effect test we collected the spleen and liver of mice to perform pathologic examinat... |
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