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The frequency and distribution of B and T lymphocyte subsets have been determined in many body tissues and fluids by preparing cell suspensions and tissue sections from lymphoid and nonlymphoid organs. In humans these studies often concentrate on the blood or on one particular cell source for obvious reasons. However, such data can only be interpreted correctly if the whole immune system is taken into consideration [64]. To facilitate this, reports on the frequencies and the absolute numbers of B and T lymphocyte subsets within various human tissues and fluids have been collected from a wide variety of journals and are briefly summarized here. Since the size of lymphoid organs varies with age (e.g. thymus, tonsils), only the data of adult individuals were included, unless otherwise stated. Natural killer (NK) cells are morphologically quite similar to lymphocytes [59], but very different functionally. For example, they are not able to recirculate from the blood via the lymph nodes and the thoracic duct back to the blood as lymphocytes do [19]. Thus, human NK cells have been compared with lymphocytes with respect to number and distribution.

Though it has been shown that immunological functions of CD4+ T cells are time of day-dependent, the underlying molecular mechanisms remain largely obscure. To address the question whether T cells themselves harbor a functional clock driving circadian rhythms of immune function, we analyzed clock gene expression by qPCR in unstimulated CD4+ T cells and immune responses of PMA/ionomycin stimulated CD4+ T cells by FACS analysis purified from blood of healthy subjects at different time points throughout the day. Molecular clock as well as immune function was further analyzed in unstimulated T cells which were cultured in serum-free medium with circadian clock reporter systems. We found robust rhythms of clock gene expression as well as, after stimulation, IL-2, IL-4, IFN-γ production and CD40L expression in freshly isolated CD4+ T cells. Further analysis of IFN-γ and CD40L in cultivated T cells revealed that these parameters remain rhythmic in vitro. Moreover, circadian luciferase reporter activity in CD4+ T cells and in thymic sections from PER2::LUCIFERASE reporter mice suggest that endogenous T cell clock rhythms are self-sustained under constant culture conditions. Microarray analysis of stimulated CD4+ T cell cultures revealed regulation of the NF-κB pathway as a candidate mechanism mediating circadian immune responses. Collectively, these data demonstrate for the first time that CD4+ T cell responses are regulated by an intrinsic cellular circadian oscillator capable of driving rhythmic CD4+ T cell immune responses.