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. 2022 Sep 24;13:5603. doi: 10.1038/s41467-022-33379-6

Fig. 5. Transmission-blocking antibodies target the N-terminal and central domains of Pfs48/45.

Fig. 5

A–C Female CD1 mice (n = 6 mice) were immunised twice with either 0.1 μg or 1 μg of Pfs48/45-D1 + 2, or 1 μg of Pfs25. Sera was collected three weeks after the final dose for analysis. A IgG titres as measured by endpoint ELISA using Pfs48/45-D1 + 2 as the coating antigen. Each symbol is for the serum sample from an individual mouse; lines represent the median of each group. Mann–Whitney two-tailed test was performed to compare the two Pfs48/45-D1 + 2 groups (p = 0.0043). B, C Transmission-blocking efficacy of IgG induced by immunisation with Pfs48/45-D1 + 2. Total IgG was purified from the pooled serum of each group (3 weeks post-boost) and mixed with P. falciparum NF54 cultured gametocytes at 750 μg/mL (B) and in a separate experiment at 375 μg/mL and 83 μg/mL (C) and fed to A. stephensi mosquitoes (n = 20 mosquitoes per test group of pooled IgG in a single feed experiment). IgG from naive mice was used as a negative control (“normal mouse Ab”); the transmission-blocking anti-Pfs25 mAb 4B7 was used as a positive control at a concentration of 94 μg/mL. Data points represent the number of oocysts in individual mosquitoes 8 days post-feeding; horizontal lines show the arithmetic mean. D, E Female CD1 mice were immunised three times with 5 μg of Pfs48/45-FL. Three weeks after the final dose sera were collected and total IgG purified. Total IgG was depleted of Pfs48/45-D3-targeting IgG using a column coupled with Pfs48/45-D3, resulting in Pfs48/45-ΔD3. D Pfs48/45-D3 specific ELISA showing lack of recognition of Pfs48/45-D3 by Pfs48/45-ΔD3. Pfs25-specific IgG (αPfs25) was included as a negative control. Data presented as mean +/− standard deviation. E Transmission-blocking efficacy of total IgG from Pfs48/45-FL or Pfs48/45-ΔD3 fed to A. stephensi mosquitoes at concentrations of 750 μg/mL, 250 μg/mL and 83 μg/mL (n = 20 mosquitoes per test group of pooled IgG in a single feed experiment). IgG from naive mice was used as a negative control (“normal mouse Ab”); transmission-blocking anti-Pfs25 mAb 4B7 was used as a positive control. Data points represent the number of oocysts in individual mosquitoes; lines show the arithmetic mean (TRA, transmission reducing activity [% inhibition in mean oocyst count per mosquito]).