Figure 3.

TD Induction of Early GC B Cells and PCs

(A)–(F) Effects of CD4⁺ T cell depletion on VRC01^gHL B cell responses to eOD-GT5 60-mer. (A) Division profiles. The gates show undivided and divided cells. (B) Divided VRC01^gHL B cell number, normalized to the 60-mer control group. (C) Undivided VRC01^gHL B cells as a frequency of total B cells, normalized to the unimmunized control. Data in (A)–(C) are from four experiments with n = 6–11 mice. Bars represent the mean. (D) Bcl6 and IRF4 expression by divided VRC01^gHL (red) and endogenous (gray) B cells. Numbers in quadrants represent percentages. (E) Divided VRC01^gHL B cells that are Bcl6^hi IRF4^lo GC B cells. (F) Divided VRC01^gHL B cells that are IRF4^hi Bcl6^lo PCs. Data in (D)–(F) are from two experiments with n = 5–6 mice. Bars represent the mean.

(G)–(L) VRC01^gHL B cell proliferation and differentiation induced by eOD-GT5_gp61 4-mer or 60-mer at day 4. (G) Division profiles. The gates show undivided and divided cells. (H) Divided VRC01^gHL B cell number. Bars represent the geometric mean. (I) Undivided VRC01^gHL B cells as a frequency of total B cells, normalized to the unimmunized control. Bars represent the mean. (J) Bcl6 and IRF4 expression in divided VRC01^gHL B cells after immunizations (blue or red) and undivided VRC01^gHL B cells from naive mice (gray). Percentages are indicated in quadrants. (K) Divided VRC01^gHL B cells that are Bcl6^hi IRF4^lo GC B cells. (L) Divided VRC01^gHL B cells that are IRF4^hi Bcl6^lo PCs. Data in (G)–(L) are from two experiments with n = 3–5 mice.

(M) Divided VRC01^gHL B cells that are Bcl6^hi early GC B cells (purple), IRF4^hi PCs (green), Bc6^hi IRF4^hi cells (black), or double negative (white). “d0” represents undivided VRC01^gHL B cells from naive mice. Data are from two to four experiments with n = 2–8 mice.

^∗∗p < 0.01; ^∗∗∗∗p < 0.0001. See also Figure S3.