Figure 1. Sustained delivery immunization enhances B_GC cell responses.

(A) Immunization and sampling schedule of first immunization. Bolus Group 2 (Gp2), 2w osmotic pump (OP), and 4w OP RMs were immunized and sampled at the same time. Bolus Gp1 were immunized and sampled separately. Bolus Gp1&2 data have been pooled.

(B) Representative B_GC cell flow cytometry, gated on viable CD20⁺ B cells pre- and post-immunization. See Fig S1 for full gating strategy.

(C) B_GC cell frequencies over time. Black circles, pooled bolus Gp1&2; grey circles, bolus Gp2.

(D) Cumulative B_GC cell responses (AUC of C) to immunization within individual LNs at weeks 1, 3–7 [AUC].

(E) Representative flow cytometry of Env trimer-specific B cells pre- and post-immunization.

(F) Env-specific B cell frequencies over time.

(G) Cumulative Env trimer-specific B cells (AUC of F) within each LN.

(H) Representative flow cytometry Env trimer-specific B_GC cells pre- and post-immunization.

(I) Env trimer-specific B_GC cell frequencies over time.

(J) Cumulative Env trimer-specific B_GC cells (AUC of I) within each LN.

(K) Env trimer-specific B_GC cells over time.

(L) Cumulative Env trimer-specific B_GC cells (AUC of K) within each LN.

(M) Frequencies of high-affinity Env trimer-specific B_GC cells over time.

(N) Cumulative high-affinity Env trimer-specific B_GC cells (AUC of M) within each LN.

(O) High-affinity Env trimer-specific B_Mem cells over time.

(P) Cumulative high-affinity Env trimer-specific B_Mem cell responses within individual LNs.

(Q) Representative flow cytometry of GC-T_FH cells, gated on CD4⁺ T cells. See Fig S2 for full gating strategy.

(R) GC-T_FH cells over time.

(S) Cumulative GC-T_FH cell response between w1 plus w3-6 [AUC].

Mean ± SEM are graphed. Statistical significance tested using unpaired, two-tailed Mann-Whitney U tests. *p≤0.05, **p≤0.01. ***p≤0.001, ****p≤0.0001

See also Figures S1–2 and Data S1.