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Immunization of mice with mycobacterial culture filtrate proteins
R D Hubbard1, C M Flory, F M Collins
1Trudeau Institute, Inc., Saranac Lake, NY 12983.
Abstract:
Culture filtrate proteins were obtained from Mycobacterium tuberculosis cultures after 7 days growth in Proskauer and Beck medium. The protein yield increased substantially to peak about the time the number of viable organisms reached its maximum level (day 8). Examination of the protein concentrate by SDS-PAGE revealed the presence of at least 12 separate protein bands varying from 10 to 90 kD. Mice were injected subcutaneously with 20 micrograms of M. tuberculosis culture filtrate (MTCF) protein suspended in saline or Freund's complete or incomplete adjuvant. The vaccinated mice were subjected to an aerogenic challenge with 10(3) colony-forming unit (CFU) M. tuberculosis Erdman and a significant reduction in the number of viable organisms was observed in the spleens and lungs determined over a 21-day period compared with age-matched normal controls. Mice immunized with the same culture filtrate proteins bound to nitrocellulose particles also showed some resistance to the virulent challenge, suggesting that individual antigens present in the culture filtrate were able to induce a protective T cell-mediated immune response in appropriately immunized mice.
Insights
Mycobacterium tuberculosis culture filtrate proteins were tested as a vaccine against tuberculosis. Immunization with these proteins significantly reduced viable bacteria in mice lungs and spleens after challenge.
Area of Science:
- Immunology
- Microbiology
- Biochemistry
Background:
- Tuberculosis (TB) remains a significant global health threat, caused by Mycobacterium tuberculosis.
- Developing effective vaccines against TB is crucial for disease control.
- Culture filtrate proteins (CFPs) from M. tuberculosis are potential vaccine candidates.
Purpose of the Study:
- To evaluate the immunoprotective potential of M. tuberculosis culture filtrate proteins (MTCF) in a mouse model.
- To investigate the immune response induced by MTCF vaccination.
Main Methods:
- M. tuberculosis CFPs were isolated and characterized using SDS-PAGE.
- Mice were immunized with MTCF proteins formulated with or without adjuvants or adsorbed onto nitrocellulose particles.
- Immunized mice were challenged aerogenically with virulent M. tuberculosis Erdman.
- Bacterial load in spleens and lungs was quantified over 21 days.
Main Results:
- SDS-PAGE analysis identified at least 12 protein bands in MTCF, ranging from 10 to 90 kDa.
- Mice vaccinated with MTCF proteins showed a significant reduction in viable M. tuberculosis in spleen and lung tissues compared to controls.
- Immunization with MTCF proteins adsorbed onto nitrocellulose particles also conferred partial protection.
Conclusions:
- M. tuberculosis culture filtrate proteins can induce a protective immune response against M. tuberculosis challenge in mice.
- The protective immunity appears to be mediated by T cell responses to specific antigens within the MTCF.
- MTCF proteins represent a promising component for developing novel TB vaccines.