Culture media selectively ensure long-term viability of <i>S</i>. <i>mansoni</i> NTS without serum or cell supplementation.

NTS were cultured in HM, M199, DMEM or RPMI supplemented with 200 U/ml Penicillin and 200 μg/ml Streptomycin for 4 weeks. (A) The viability of the parasites was scored at the indicated time points and (B) the morphology was observed under light microscopy. Photomicrographs were taken 2 and 4 weeks p.t. Scale bar applies to all shown images. Results are representative of at least three independent experiments. Each data point is shown as mean ± SD of at least three biological replicates. p.t., post-transformation; s.p., score points; HM, HybridoMed Diff 1000. (PZFX)</p

Drug sensitivity is dependent on the developmental stage of <i>S</i>. <i>mansoni</i> larvae generated <i>in vitro</i>.

NTS were cultured and matured in HM supplemented with 200 U/ml Penicillin and 200 μg/ml Streptomycin only or additionally supplemented with 20% HSe. 100, 10 and 1 μg/ml of (A) praziquantel, (B) oxamniquine, (C) mefloquine and (D) artemether were then added to the culture 1 day p.t. to test the SkS, 7 days p.t. to test the LuS and 6 weeks p.t. to test the LiS. Viability was scored 72h a.t. Each data point is shown as a mean ± SD of three independent experiments with at least three biological replicates each. ××× p ≤ 0.001, ×× p ≤ 0.01, × p ≤ 0.05 comparing control with 1 μg/ml drug; ┴┴┴ p ≤ 0.001, ┴┴ p ≤ 0.01, ┴ p ≤ 0.05 comparing control with 10 μg/ml drug; ***p ≤ 0.001, **p ≤ 0.01 *p ≤ 0.05 control with 100 μg/ml drug. SkS, skin stage; LuS, lung stage; LiS, liver stage; PZQ, praziquantel; OXM, oxamniquine; MFQ, mefloquine; ART, artemether; s.p., score points. (PZFX)</p

Presence of human serum <i>in vitro</i> enhances NTS viability and induces worm development past the LuS.

NTS were cultured in HM and DMEM supplemented with 200 U/ml Penicillin and 200 μg/ml Streptomycin with or without additional supplementation of 20% HSe. (A) Viability scoring was performed at the indicated time points. The percentage of developmental stages in culture with either (B) HM, (C) HM + 20% HSe, (D) DMEM or (E) DMEM + 20% HSe was calculated for the indicated time points by bright field microscopy. (F) Representative photomicrographs were taken on day 35 post-transformation. Scale bar applies to all shown pictures. Arrowheads indicate dead NTS. Arrows indicate early and late LiS. Each data point is shown as a mean ± SD of pooled data from three independent experiments with three biological replicates each. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 comparing HM with HM + 20% HSe, × p ≤ 0.05, ×× ≤ 0.01, ××× ≤ 0.001 comparing DMEM with DMEM + 20%. HSe, human serum; SkS, skin stage; LuS, lung stage; LiS, liver stage; s.p., score points; p.t., post-transformation. (PZFX)</p

Efficient drug screening of advanced larval stages of <i>S</i>. <i>mansoni</i> generated in human serum.

NTS were cultured in HM supplemented with 200 U/ml Penicillin and 200 μg/ml Streptomycin and 20% HSe for 1 week to generate LuS (B, D, F and H) and 6 weeks to obtain LiS (A, C, E, G) schistosomula. (A, B) Praziquantel (PZQ), (C, D) Oxamniquine (OXM), (E, F) Mefloquine (MFQ) and (G, H) Artemether (ART) were dissolved in DMSO and added at indicated concentrations for the entire duration of the experiment. 1% DMSO in culture medium served as control. Viability was scored before treatment (0 h) and 3 hours, 1, 2, 3 and 7 days a.t. Each data point is shown as a mean ± SD of three independent experiments with at least three biological replicates each. ××× p ≤ 0.001, ×× p ≤ 0.01, × p ≤ 0.05 comparing control with 1 μg/ml drug; ┴┴┴ p ≤ 0.001 ┴┴ p ≤ 0.01 ┴ p ≤ 0.05 control with 10 μg/ml drug; ***p ≤ 0.001, **p ≤ 0.01 *p ≤ 0.05 control with 100 μg/ml drug. LuS, lung stage; LiS, liver stage; s.p., score points; a.t., after treatment. (PZFX)</p

HSe supplemented HM increased the development to late liver stage compared to Basch medium 169.

NTS of Schistosoma mansoni (NMRI strain) were cultured in HM and Basch-Medium 169 supplemented with 200 U/ml Penicillin and 200 μg/ml Streptomycin as well as additional supplementation of either 20% FCS or 20% HSe. (A) Viability scoring was performed at the indicated time points. The percentage of developmental stages in culture with (B) Basch medium 169 supplemented with (C) 20% FCS or (D) 20% HSe as well as (E) HM supplemented with (F) 20% FCS or (G) 20% HSe were calculated per well for the indicated time points by bright field microscopy. (H) Representative photomicrographs were taken on day 28 p.t. Scale bar applies to all images shown. Arrowheads indicate dead NTS. Arrows indicate early and late LiS. Each data point is shown as a mean ± SD of an experiment with three biological replicates each. FCS, fetal calf serum; HSe, human serum; SkS, skin stage; LuS, lung stage; LiS, liver stage; s.p., score points; p.t., post-transformation. (PZFX)</p

Generation of late LiS worm in medium supplemented with human serum increases in a concentration-dependent manner.

NTS were cultured in HM supplemented with 200 U/ml Penicillin and 200 μg/ml Streptomycin as well as 1%, 5%, 10%, 20% and 50% of HSe. The percentages of the developmental stages as well as dead parasites in culture with (A) HM alone or supplemented with (B) 1%, (C) 20% and (D) 50% HSe were calculated at indicated time points and (E) their viability was scored during bright field microscopy. (F) Representative photomicrographs were taken on day 35 post transformation. Scale bar applies to all pictures shown. Arrowheads indicate dead NTS. Arrows indicate LiS (early LiS in 5% and 10% HSe) and late liver stage (in 20% and 50% HSe). Results are pooled from three independent experiments with at least three biological replicates each. ×× p ≤ 0.01 comparing HM vs. 1% HSe; ┴┴ p ≤ 0.01 comparing HM vs. 5% HSe; ˅˅ p ≤ 0.01 comparing HM vs. 10% HSe; ≠≠ p ≤ 0.01 comparing HM vs. 20% HSe; ** comparing p ≤ 0.01 HM vs. 50% HSe. HSe, human serum; s.p., score points; p.t., post-transformation; SkS, skin stage; LuS, lung stage; LiS, liver stage. (PZFX)</p

Image_1_Praziquantel Reduces Maternal Mortality and Offspring Morbidity by Enhancing Anti-Helminthic Immune Responses.tiff

Alongside the wide distribution throughout sub Saharan Africa of schistosomiasis, the morbidity associated with this chronic parasitic disease in endemic regions is often coupled with infection-driven immunomodulatory processes which modify inflammatory responses. Early life parasite exposure is theorized to drive immune tolerance towards cognate infection as well as bystander immune responses, beginning with in utero exposure to maternal infection. Considering that 40 million women of childbearing-age are at risk of infection worldwide, treatment with Praziquantel during pregnancy as currently recommended by WHO could have significant impact on disease outcomes in these populations. Here, we describe the effects of anthelminthic treatment on parasite-induced changes to fetomaternal cross talk in a murine model of maternal schistosomiasis. Praziquantel administration immediately prior to mating lead to clear re-awakening of maternal anti-parasite immune responses, with persistent maternal immune activation that included enhanced anti-schistosome cytokine responses. Clearance of parasites also improved capacity of dams to endure the additional pressure of pregnancy during infection. Maternal treatment also drove lasting functional alterations to immune system development of exposed offspring. Prenatal anthelminthic treatment skewed offspring immune responses towards parasite clearance and reduced morbidity during cognate infection. Maternal treatment also restored offspring protective IgE antibody responses directed against schistosome antigens, which were otherwise suppressed following exposure to untreated maternal infection. This was further associated with enhanced anti-schistosome cytokine responses from treatment-exposed offspring during infection. In the absence of cognate infection, exposed offspring further demonstrated imprinting across cellular populations. We provide further evidence that maternal treatment can restore a more normalized immune profile to such offspring exposed in utero to parasite infection, particularly in B cell populations, which may underlie improved responsiveness to cognate infection, and support the WHO recommendation of anthelminthic treatment during pregnancy.</p

Image_4_Praziquantel Reduces Maternal Mortality and Offspring Morbidity by Enhancing Anti-Helminthic Immune Responses.tiff

Alongside the wide distribution throughout sub Saharan Africa of schistosomiasis, the morbidity associated with this chronic parasitic disease in endemic regions is often coupled with infection-driven immunomodulatory processes which modify inflammatory responses. Early life parasite exposure is theorized to drive immune tolerance towards cognate infection as well as bystander immune responses, beginning with in utero exposure to maternal infection. Considering that 40 million women of childbearing-age are at risk of infection worldwide, treatment with Praziquantel during pregnancy as currently recommended by WHO could have significant impact on disease outcomes in these populations. Here, we describe the effects of anthelminthic treatment on parasite-induced changes to fetomaternal cross talk in a murine model of maternal schistosomiasis. Praziquantel administration immediately prior to mating lead to clear re-awakening of maternal anti-parasite immune responses, with persistent maternal immune activation that included enhanced anti-schistosome cytokine responses. Clearance of parasites also improved capacity of dams to endure the additional pressure of pregnancy during infection. Maternal treatment also drove lasting functional alterations to immune system development of exposed offspring. Prenatal anthelminthic treatment skewed offspring immune responses towards parasite clearance and reduced morbidity during cognate infection. Maternal treatment also restored offspring protective IgE antibody responses directed against schistosome antigens, which were otherwise suppressed following exposure to untreated maternal infection. This was further associated with enhanced anti-schistosome cytokine responses from treatment-exposed offspring during infection. In the absence of cognate infection, exposed offspring further demonstrated imprinting across cellular populations. We provide further evidence that maternal treatment can restore a more normalized immune profile to such offspring exposed in utero to parasite infection, particularly in B cell populations, which may underlie improved responsiveness to cognate infection, and support the WHO recommendation of anthelminthic treatment during pregnancy.</p

Image_3_Praziquantel Reduces Maternal Mortality and Offspring Morbidity by Enhancing Anti-Helminthic Immune Responses.tiff

Alongside the wide distribution throughout sub Saharan Africa of schistosomiasis, the morbidity associated with this chronic parasitic disease in endemic regions is often coupled with infection-driven immunomodulatory processes which modify inflammatory responses. Early life parasite exposure is theorized to drive immune tolerance towards cognate infection as well as bystander immune responses, beginning with in utero exposure to maternal infection. Considering that 40 million women of childbearing-age are at risk of infection worldwide, treatment with Praziquantel during pregnancy as currently recommended by WHO could have significant impact on disease outcomes in these populations. Here, we describe the effects of anthelminthic treatment on parasite-induced changes to fetomaternal cross talk in a murine model of maternal schistosomiasis. Praziquantel administration immediately prior to mating lead to clear re-awakening of maternal anti-parasite immune responses, with persistent maternal immune activation that included enhanced anti-schistosome cytokine responses. Clearance of parasites also improved capacity of dams to endure the additional pressure of pregnancy during infection. Maternal treatment also drove lasting functional alterations to immune system development of exposed offspring. Prenatal anthelminthic treatment skewed offspring immune responses towards parasite clearance and reduced morbidity during cognate infection. Maternal treatment also restored offspring protective IgE antibody responses directed against schistosome antigens, which were otherwise suppressed following exposure to untreated maternal infection. This was further associated with enhanced anti-schistosome cytokine responses from treatment-exposed offspring during infection. In the absence of cognate infection, exposed offspring further demonstrated imprinting across cellular populations. We provide further evidence that maternal treatment can restore a more normalized immune profile to such offspring exposed in utero to parasite infection, particularly in B cell populations, which may underlie improved responsiveness to cognate infection, and support the WHO recommendation of anthelminthic treatment during pregnancy.</p

Image_2_Praziquantel Reduces Maternal Mortality and Offspring Morbidity by Enhancing Anti-Helminthic Immune Responses.tiff

Alongside the wide distribution throughout sub Saharan Africa of schistosomiasis, the morbidity associated with this chronic parasitic disease in endemic regions is often coupled with infection-driven immunomodulatory processes which modify inflammatory responses. Early life parasite exposure is theorized to drive immune tolerance towards cognate infection as well as bystander immune responses, beginning with in utero exposure to maternal infection. Considering that 40 million women of childbearing-age are at risk of infection worldwide, treatment with Praziquantel during pregnancy as currently recommended by WHO could have significant impact on disease outcomes in these populations. Here, we describe the effects of anthelminthic treatment on parasite-induced changes to fetomaternal cross talk in a murine model of maternal schistosomiasis. Praziquantel administration immediately prior to mating lead to clear re-awakening of maternal anti-parasite immune responses, with persistent maternal immune activation that included enhanced anti-schistosome cytokine responses. Clearance of parasites also improved capacity of dams to endure the additional pressure of pregnancy during infection. Maternal treatment also drove lasting functional alterations to immune system development of exposed offspring. Prenatal anthelminthic treatment skewed offspring immune responses towards parasite clearance and reduced morbidity during cognate infection. Maternal treatment also restored offspring protective IgE antibody responses directed against schistosome antigens, which were otherwise suppressed following exposure to untreated maternal infection. This was further associated with enhanced anti-schistosome cytokine responses from treatment-exposed offspring during infection. In the absence of cognate infection, exposed offspring further demonstrated imprinting across cellular populations. We provide further evidence that maternal treatment can restore a more normalized immune profile to such offspring exposed in utero to parasite infection, particularly in B cell populations, which may underlie improved responsiveness to cognate infection, and support the WHO recommendation of anthelminthic treatment during pregnancy.</p