V beta 6+ and V beta 4+ T cells exert cooperative activity in clearance of secondary infection with Histoplasma capsulatum.

We previously studied the lung Vbeta TCR repertoire of C57BL/6 mice during primary infection with the pathogen Histoplasma capsulatum. We observed a consistent oligoclonal expansion of Vbeta4(+) T cells during the peak of infection and early stages of resolution. The Vbeta4(+) family played a role in protective immunity against the fungus. Depletion of this subpopulation of T cells hindered optimal clearance of infection from tissues. In this report we analyze the flux of the Vbeta TCR repertoire in the lungs of C57BL/6 mice with reinfection histoplasmosis. We observed a significant increase in Vbeta6(+) T cells on days 7, 10, and 14, the peak and early resolution phases of infection. This skewing was preceded by an increased number of memory T cells within Vbeta6(+) cells. The VDJ sequences of Vbeta6 chains were oligoclonal during the early stages of the infection, suggesting that the expansion was driven by a small number of Ags. More than 96% of the expanded Vbeta6(+) cells were CD4(+). Depletion of Vbeta6(+) T cells but not Vbeta4(+) T cells induced a modest but significant delay in fungal clearance. Simultaneous depletion of Vbeta4(+) and Vbeta6(+) T cells induced a more pronounced impairment of host resistance. These studies illustrate the dynamic interactions between Vbeta families in the response to microbial challenge.

Predominant usage of the proximal poly(A) site in alpha mRNAs is not intrinsic to the 3' termini.

The maturation of IgM-expressing B cells to IgM-secreting plasma cells is associated with both an increase in mu mRNA and the ratio of secreted to membrane forms of mu mRNA. In contrast, previous studies demonstrated that in vitro the secreted form of alpha mRNA (alpha s mRNA) predominates regardless of the stage of B cell differentiation. The present study demonstrates that alpha s mRNA predominates in both B cells derived from the germinal centers of murine Peyer's patches and in the functional IgA memory population, suggesting that in vitro events accurately represent the generation of a secretory IgA response in vivo. Although the predominant usage of the alpha s poly(A) site is due to RNA processing, it does not depend on either the alpha s poly(A) site, the 3' splice site associated with the exon encoding the membrane exon of IgA (alphaM) or the alphaM poly(A) sites. Analysis of the sequence of the intron between the alpha s terminus and alphaM (alpha s-alphaM intron) demonstrates the existence of several potential regulatory elements. Furthermore, the effects of deletions within the alpha s-alphaM intron on 3' terminus usage demonstrate that the predominant usage of the proximal terminus is not strictly dependent on the length of the intron. Together with previous work, these observations support the idea that choice of 3' terminus for all Ig heavy chain genes is regulated by a similar mechanism, but specific sequences within a heavy chain gene can impinge upon that mechanism.