Persistence and enrichment of dominant T cell clonotypes in expanded tumor-infiltrating lymphocytes of breast cancer.

BACKGROUND: Adoptive cell therapy using tumor-infiltrating lymphocytes (TILs) has shown promising results in cancer treatment, including breast cancer. However, clonal dynamics and clinical significance of TIL expansion ex vivo remain poorly understood. METHODS: We investigated T cell receptor (TCR) repertoire changes in expanded TILs from 19 patients with breast cancer. We compared TCR repertoire of TILs at different stages of expansion, including initial (2W TILs) and rapid expansion (REP TILs), and their overlap with formalin fixed paraffin embedded (FFPE) and peripheral blood. Additionally, we examined differences in TCR repertoire between CD4+ and CD8+ REP TILs. RESULTS: In descending order of proportion, average of 60% of the top 10% clonotypes of FFPE was retained in 2W TIL (60% in TRB, 64.7% in TRA). Among the overlapped clonotypes between 2W TILs and REP TILs, 69.9% was placed in top 30% of 2W TIL. The proportion of clonotypes in 2W TIL and REP TIL showed a significant positive correlation. CD4+ and CD8+ T cells show similar results in diversity and CDR3 length. CONCLUSIONS: Our study traces the changes in TILs repertoire from FFPE to 2W TIL and REP TIL and confirmed that clonotypes with high frequencies in TILs have a high likelihood of maintaining their priority throughout culture process.

Dependence of transverse and longitudinal resolutions on incident Gaussian beam widths in the illumination part of optical scanning microscopy.

We studied both theoretically and experimentally the intensity distribution of a Gaussian laser beam when it was focused by an objective lens with its numerical aperture up to 0.95. Approximate formulas for full width at half-maximum (FWHM) of the intensity distribution at focus were derived for very large and very small initial beam waists with respect to the entrance pupil radius of the objective lens. In experiments, the energy flux through a 0.5 microm pinhole was measured for various pinhole positions. It was found in theoretical analysis and confirmed in experiments that the FWHMs at focus in the transverse and longitudinal directions do not increase much from the ultimate FWHMs until the input beam waist is reduced below half of the entrance pupil radius.