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. 2022 Jun 13;2(6):100238. doi: 10.1016/j.crmeth.2022.100238

Figure 2.

Figure 2

Identification of latently and productively infected CD4+ T cells

(A) Experimental design to analyze CD4+ T cells infected with pMorpheus-V5. CD4+ T cells from four healthy donors were stimulated with IL-2 or a combination of IL-2 with α-CD3/CD28 antibody-coated beads for 3 days prior to infection with pMorpheus-V5. The frequency of latently and productively infected cells was determined 5 dpi using flow cytometry.

(B) Frequency of latently (V5POS, NGFRPOS; yellow) and productively (mCherryPOS, V5POS, NGFRPOS, p24POS; green) infected CD4+ T cells stimulated with IL-2 alone or IL-2/α-CD3/CD28. Cells were analyzed using flow cytometry 5 dpi. Each of the four healthy donors is identified by symbol. Error bars represent the average ± SD, and statistical significance was established using unpaired t test.

(C) Relative proportion of pMorpheus-V5 latently (yellow) or productively (green) infected CD4+ T cells stimulated with IL-2 is shown. The data represent the average of the four healthy donors. Average LTR-silent 58% ± 4% versus average LTR-active 42% ± 4% (p = 0.0014, unpaired t test).

(D) Relative proportion of pMorpheus-V5 latently (yellow) or productively (green) infected CD4+ T cells activated with α-CD3/CD28. The data represent the average of the four individual healthy donors. Average LTR-silent 52% ± 8% versus average LTR-active 48% ± 8% (p = 0.61, t test).