Tissue Trafficking Kinetics of Rhesus Macaque Natural Killer Cells Measured by Serial Intravascular Staining

Abstract

The in vivo tissue distribution and trafficking patterns of natural killer (NK) cells remain understudied. Animal models can help bridge the gap, and rhesus macaque (RM) primates faithfully recapitulate key elements of human NK cell biology. Here, we profiled the tissue distribution and localization patterns of three NK cell subsets across various RM tissues. We utilized serial intravascular staining (SIVS) to investigate the tissue trafficking kinetics at steady state and during recovery from CD16 depletion. We found that at steady state, CD16+ NK cells were selectively retained in the vasculature while CD56+ NK cells had a shorter residence time in peripheral blood. We also found that different subsets of NK cells had distinct trafficking kinetics to and from the lymph node as well as other lymphoid and non-lymphoid tissues. Lastly, we found that following administration of CD16-depleting antibody, CD16+ NK cells and their putative precursors retained a high proportion of continuously circulating cells, suggesting that regeneration of the CD16 NK compartment may take place in peripheral blood or the perivascular compartments of tissues.

Barcode clonal tracking of tissue-resident immune cells in rhesus macaque highlights distinct clonal distribution pattern of tissue NK cells

“…Therefore, partial overlap between NK clones observed in PB and tissue may be from PB contamination, especially clones detected in the spleen and liver. However, our previous studies using intravascular staining to follow immune cell dynamics in macaques ( 42 , 76 ) found very limited circulating CD45+ hematopoietic cells contaminating gut and LN biopsies. Our phenotypic analyses of NK cells in different tissues and PB revealed marked differences, even for liver or spleen, indicating the cells we obtained from all tissues have limited PB contamination.…”

“…We demonstrated groups of balanced/un-biased CD56+ NK clones and DN NK clones contributing equivalently in PB CD56+ NK compared to tissues, suggesting a shared clonal repertoire between those populations. We recently utilized serial intravascular staining (SIVS) with multi-color labeled CD45 antibodies immediately binding to circulating blood cells to study movement of blood immune cells in and out of tissues ( 42 , 76 ). We found that PB CD56+ and DN NK cells have a shorter residence time in blood and traffic more often between PB and tissues such as lymph nodes compared to the CD16+ NK subset, which remains primarily within the circulation.…”

“…The available human anti-NKG2A antibodies recognize both activating NKG2C and inhibitory NKG2A receptor isoforms ( 37 , 38 , 40 ). Based on CD16 and CD56 expression, NKG2A/C+ NK cells in RM are classified into three distinct NK subsets: CD16+CD56- (CD16+), CD56+CD16- (CD56+) and CD56-CD16- (double negative or “DN”) ( 41 , 42 ). RM CD56+ and CD16+ NK cells closely resemble human CD56 bright and CD56 dim NK cells, respectively ( 38 , 43 , 44 ).…”

“…The ontogeny of these human intermediate subsets is not well established. Using intravascular staining, we recently reported distinct trafficking dynamics for each RM NK cell population between blood and tissues ( 42 ).…”

Lingering Effects of Early Institutional Rearing and Cytomegalovirus Infection on the Natural Killer Cell Repertoire of Adopted Adolescents

“…The lower percent of CD56 bright NK cells likely reflects a shift in the distribution and trafficking patterns of NK cells, with more CD56 bright NK cells remaining resident in tissue [71][72][73][74]. This NK cell subset can also egress from the vasculature more readily than other NK cells and return back to tissue, including to lymph nodes and the liver.…”