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Some of the proteins of mycobacteria are preferentially associated with the cell wall and are powerful immunogens, and humoral antibody responses to these mycobacterial antigens may occur in patients with tuberculosis. In this study, Triton X-100 solubilized protein (TSP) antigen was isolated from Mycobacterium tuberculosis H37Rv by overnight shaking with 1% Triton X- 100/PMSF and 10-90% ammonium sulfate precipitation. IgG and IgM antibody levels against TSP, crude protein from the unheated cultrue filtrate (CF#) and 30 kDa antigens were determined in the sera of 80 patients with pulmonary tuberculosis and 99 healthy controls with PPD (+) and (-). High IgG reactivity to TSP and CF antigen was observed in tuberculosis patients. Mean IgG antibody titers against all of three mycobacterial antigens were differed significantly (P<0.01) between patients and controls but IgM showed no difference. By the cut-off value adding 2 standard deviation to the mean absorbance of controls, the sensitivity and specificity of the IgG antibody to TSP antigen were 93.9% and 77.5%. The specificity to TSP antigen was a litttle higher than those obtained by CF and 30 kDa antigen. From the above results, the TSP antigen may be useful for the serodiagnosis of tuberculosis.
Ammonium Sulfate ; Antibody Formation ; Cell Wall ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Mycobacterium tuberculosis* ; Mycobacterium* ; Neptune* ; Octoxynol* ; Sensitivity and Specificity ; Serologic Tests ; Tuberculosis ; Tuberculosis, Pulmonary*

10.In vitro Stimulation of Tumor - Draining Lymph Node Lymphocytes with the 30 kDa Antigen of Mycobacterium tuberculosis Leads to the Differentiation of Th1 Cells and Cytotoxic Effector Cells.

Jeong Kyu PARK ; Tae Hyun PAIK ; Seok Shin KOH ; Hwa Jung KIM ; Eun Kyeong JO

Korean Journal of Immunology 1997;19(1):59-72

Tumor-draining lymph node (TDLN) lymphocytes contain immunologically sensitized to tumor but functionally deficient T cells. The 30 kDa protein antigen, a major secreted protein antigen of Mycobacterium tuberculosis, exhibits strong T cell stimulatory effect. In this study, it examined that the feasibility of using M tuberculosis 30 kDa antigen to stimulate tumor-draining lymph node cells for the generation of specific immune effector cells. Freshly isolated TDLN lymphocytes could directly respond to the 30 kDa antigen alone and their proliferative responses were markedly augmented by stimulation with rIL-2. TDLN cells were stimulated with the 30 kDa antigen for various time intervals and examined for the induction of IFN-r and IL-4 mRNA using RT-PCR. The expression of IFN-r mRNA was greatly augmented after 1 wk, whereas IL-4 mRNA is markedly decreased after 1 wk. Cytotoxic T cell activities induced by the 30 kDa antigen was also evaluated. TDLN cells stimulated with the 30 kDa antigen alone were able to generate remarkable cytotoxic response to K562 or Daudi cell lines after 6 days of culture. And their cytotoxic effects were highly augmented by stirnulation with rIL-2. These results suggest that the 30 kDa antigen of M. tuberculosis may selectively activate Thl cells of TDLN lymhocytes and induce the cytotoxic T cell activities. In conclusion, the 30 kDa antigen can be used as a biologic response modifier in tumor immunology.
Allergy and Immunology ; Cell Line ; Interleukin-4 ; Lymph Nodes* ; Lymphocytes* ; Mycobacterium tuberculosis* ; Mycobacterium* ; RNA, Messenger ; T-Lymphocytes ; Th1 Cells* ; Tuberculosis

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3.Purification and immunochemical charaterization of alpha-antigen from the culture filtrate of mycobacterium tuberculosis.

4.Determination of antibody activities of alpha- and beta-protein antigens of mycobacterium tuberculosis in cerebrospinal fluid by ELISA for the diagnosis of tuberculous meningitis.

5.Lymphocyte proliferation and antibody response against 30-kDa protein antigen of mycobacterium tuberculosis.

6.Detection of mycobacterium tuberculosis in sputum samples by polymerase chain reaction.

7.Purification of 30-kDa and 32 kDa protein antigens from mycobacterium tuberculosis and activation of human monocytes by lymphokines.

8.Biological detection of enterotoxigenic E. coli.

9.Determination of Anti-tuberculous Antibody Against Triton X-100 Solubilized Protein ( TSP ) Antigen of Mycobacterium tuberculosis in the Sera of Patients with Pulmonary Tuberculosis.

10.In vitro Stimulation of Tumor - Draining Lymph Node Lymphocytes with the 30 kDa Antigen of Mycobacterium tuberculosis Leads to the Differentiation of Th1 Cells and Cytotoxic Effector Cells.

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