T-cell stimuli independently sum to regulate an inherited clonal division fate

Abstract

In the presence of antigen and costimulation, T cells undergo a characteristic response of expansion, cessation and contraction. Previous studies have revealed that population-level reproducibility is a consequence of multiple clones exhibiting considerable disparity in burst size, highlighting the requirement for single-cell information in understanding T-cell fate regulation. Here we show that individual T-cell clones resulting from controlled stimulation in vitro are strongly lineage imprinted with highly correlated expansion fates. Progeny from clonal families cease dividing in the same or adjacent generations, with inter-clonal variation producing burst-size diversity. The effects of costimulatory signals on individual clones sum together with stochastic independence; therefore, the net effect across multiple clones produces consistent, but heterogeneous population responses. These data demonstrate that substantial clonal heterogeneity arises through differences in experience of clonal progenitors, either through stochastic antigen interaction or by differences in initial receptor sensitivities.

Superior properties of CellTrace Yellow™ as a division tracking dye for human and murine lymphocytes

“…Thus, CTY can facilitate existing automated proliferation analysis tools to estimate cell numbers in each generation, and extract proliferation, survival, and differentiation information as required for many quantitative applications. 10,12,[15][16][17] During our investigation of recently available cell division tracking dyes, we determined that poor CTFR peak resolution was improved by sorting labeled cells prior to first division for a narrow fluorescence range, generating clear division peaks in subsequent analyses. Thus, poor peak resolution can be attributed to broad, inconsistent labeling of the initial cell population, presumably due to slight dye uptake differences between CTV and CTY, created by their unique chemistry.…”

“…These include the discoveries that T and B lymphocytes share a common regulatory process where changes in class of the response are linked to clonal expansion, and that cell division times are variable and stochastic, but highly concordant in families. [9][10][11][12][13][14][15][16][17] These dyes have also been used to study molecular symmetry at mitosis, and to investigate asymmetric cell division in lymphocytes. [18][19][20] CFSE has been used in several studies, alone or in combination with other dyes, to investigate cytotoxic T cell killing or regulatory T cell suppression of target cells, 6,[21][22][23][24] as well as in the investigation of NK cell proliferation regulation.…”

Modifying clonal selection theory with a probabilistic cell

“…In other situations, recently discovered, division will have no effect on the time to stop dividing or the time to die . This latter rule should lead to families that share timed events (such as division cessation or death) in the same generation, as is frequently observed by experiment . Given the mechanics and rules of interaction between components, and the effects to be altered over time, a calculation scheme to model the population cell dynamics in number and type is possible.…”

“…59 This latter rule should lead to families that share timed events (such as division cessation or death) in the same generation, 59 as is frequently observed by experiment. 50,60,61 Given the mechanics and rules of interaction between components, and the effects to be altered over time, a calculation scheme to model the population cell dynamics in number and type is possible. As processes are operating and dictating cell changes over time and require signal integration and affect differentiation, it is reminiscent of the calculus, and with Amanda Gett, we labeled these operations as the Cellular Calculus.…”

“…Evidence that these molecular "construction" differences are significant is highlighted by the finding of remarkable fate similarities between siblings and families in vitro and even in vivo. [50][51][52]60,61,83,84 It appears if cells are molecular "clones" they behave almost identically, and that the large amount of diversity between similar cells might presumably be traced to a set of molecular expression differences.…”