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Infection and Immunity, October 1998, p. 4981-4988, Vol. 66, No. 10
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

An Ex Vivo Study of T Lymphocytes Recovered from the Lungs of I/St Mice Infected with and Susceptible to Mycobacterium tuberculosis

Irina Lyadova,1,* Vladimir Yeremeev,1 Konstantin Majorov,1 Boris Nikonenko,1 Sergei Khaidukov,2 Tatiana Kondratieva,1 Natalia Kobets,1 and Alexander Apt1

Department of Immunology, Central Institute for Tuberculosis, Moscow 107564,1 and Laboratory for Immunochemistry, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Science, Moscow 117871,2 Russia

Received 17 February 1998/Returned for modification 1 April 1998/Accepted 2 July 1998

I/St mice, previously characterized as susceptible to Mycobacterium tuberculosis H37Rv, were given 103 or 105 CFU intravenously. At two time points postinoculation, the cell suspensions that resulted from enzymatic digestion of lungs were enumerated and further characterized phenotypically and functionally. Regarding the T-cell populations recovered at 2 and 5 weeks postinfection, two main results were obtained: (i) the population of CD44- CD45RB+ cells disappeared within 2 weeks postinfection, while the number of CD44+ CD45RB-/low cells slowly increased between weeks 2 and 5; (ii) when cocultured with irradiated syngeneic splenocytes, these lung T cells proliferated in the presence of H37Rv sonicate. Using H37Rv sonicate and irradiated syngeneic splenocytes to reactivate lung T cells, we selected five CD3+ CD4+ CD8- T-cell clones. In addition to the H37Rv sonicate, the five clones react to both a short-term culture filtrate and an affinity-purified 15- to 18-kDa mycobacterial molecule as assessed by the proliferative assay. However, there was a clear difference between T-cell clones with respect to cytokine (gamma interferon [IFN-gamma ] and interleukin-4 [IL-4] and IL-10) profiles: besides one Th1-like (IFN-gamma + IL-4-) clone and one Th0-like (IFN-gamma + IL-4+ IL-10+) clone, three clones produced predominantly IL-10, with only marginal or no IL-4 and IFN-gamma responses. Inhibition of mycobacterial growth by macrophages in the presence of T cells was studied in a coculture in vitro system. It was found that the capacity to enhance antimycobacterial activity of macrophages fully correlated with INF-gamma production by individual T-cell clones following genetically restricted recognition of infected macrophages. The possible functional significance of cytokine diversity among T-cell clones is discussed.

* Corresponding author. Mailing address: Laboratory for Immunogenetics, Central Institute for Tuberculosis, Yauza alley, 2, 107564 Moscow, Russia. Phone: (095) 268 78 10. Fax: (095) 963 80 44. E-mail: asapt{at}aha.ru.

Infection and Immunity, October 1998, p. 4981-4988, Vol. 66, No. 10
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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