Vaccine-Induced Carbohydrate-Specific Memory B Cells Reactivate During Rodent Malaria Infection

Hayley Joseph

Qiao Tan

Ramin Mazhari

Emily Eriksson

Louis Schofield

Supplementary Figure 1

Flow cytometry gating strategy. This is the flow cytometry gating strategy using an example of a spleen isolated from a GPI-immunized mouse at day 14.

Click here for additional data file.^{(1.3M, TIF)}

Supplementary Figure 2

Representative plots for experimental groups; one mouse from each group.

Click here for additional data file.^{(1.2M, TIF)}

Supplementary Figure 3

Challenge with synthetic GPI-KLH or pRBCs induced rapid activation of GPI-specific MBCs in previously GPI-vaccinated mice, whilst Plasmodium mice do not generate GPI-specific MBC. Mice were vaccinated and boosted with GPI-KLH (n = 24) or infected twice with pRBCs i.p. (n = 24, referred to as Plasmodium mice). Naïve mice were included as a control group (n = 24). After resting for a minimum of 10 weeks, mice were injected with GPI-KLH i.p. or 1 × 10^pRBCsi.p. to assess reactivation of MBCs at day 5. The supplementary figure outlines FACS plots.

Click here for additional data file.^{(995K, TIF)}

Supplementary Table 1

Vaccination/Infection protocol for assessing B cell activation kinetics and memory B cell recall.

Click here for additional data file.^{(20K, DOCX)}

^{Division of Population Health and Immunity, Walter and Eliza Hall Institute of Medical Research, Melbourne, VIC, Australia}

^{Department of Medical Biology, The University of Melbourne, Melbourne, VIC, Australia}

^{Australian Institute of Tropical Health and Medicine, James Cook University, Townsville, QLD, Australia}

Edited by: Wanderley De Souza, Federal University of Rio de Janeiro, Brazil

Reviewed by: Moriya Tsuji, Aaron Diamond AIDS Research Center, United States; Ricardo Fujiwara, Federal University of Minas Gerais, Brazil

*Correspondence: Emily M. Eriksson ua.ude.ihew@nosskire

This article was submitted to Microbial Immunology, a section of the journal Frontiers in Immunology

Edited by: Wanderley De Souza, Federal University of Rio de Janeiro, Brazil

Reviewed by: Moriya Tsuji, Aaron Diamond AIDS Research Center, United States; Ricardo Fujiwara, Federal University of Minas Gerais, Brazil

Received 2019 Jan 18; Accepted 2019 Jul 22.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

Abstract

A long-standing challenge in malaria is the limited understanding of B cell immunity, previously hampered by lack of tools to phenotype rare antigen-specific cells. Our aim was to develop a method for identifying carbohydrate-specific B cells within lymphocyte populations and to determine whether a candidate vaccine generated functional memory B cells (MBCs) that reactivated upon challenge with Plasmodium (pRBCs). To this end, a new flow cytometric probe was validated and used to determine the kinetics of B cell activation against the candidate vaccine glycosylphosphatidylinositol conjugated to Keyhole Limpet Haemocyanin (GPI-KLH). Additionally, immunized C57BL/6 mice were rested (10 weeks) and challenged with pRBCs or GPI-KLH to assess memory B cell recall against foreign antigen. We found that GPI-specific B cells were detectable in GPI-KLH vaccinated mice, but not in Plasmodium-infected mice. Additionally, in previously vaccinated mice GPI-specific IgG1 MBCs were reactivated against both pRBCs and synthetic GPI-KLH, which resulted in increased serum levels of anti-GPI IgG in both challenge approaches. Collectively our findings contribute to the understanding of B cell immunity in malaria and have important clinical implications for inclusion of carbohydrate conjugates in malaria vaccines.

Keywords: malaria, B cell immunity, vaccines, memory failure, glycosylphosphatidylinositol (GPI)

Abstract

Acknowledgments

The authors would like to thank Prof. David Tarlinton and Dr. Kim Jacobson for their invaluable input and the Bundoora Antibody facility for generating IgD-A680 and Gr-A680.

Acknowledgments

Glossary

Abbreviations

BIO	biotin labelling
BSA	bovine serum albumin
GPI	glycosylphosphatidylinositol
KLH	Keyhole Limpet Haemocyanin
MBC	memory B cells
NP	Four-hydroxy-3-nitrophenyl acetyl-Osu
pRBCs	parasitized red blood cells.

Glossary

Footnotes

Funding. This work was supported by the National Health and Medical Research Council [GNT1052580 to HJ and GNT1093311] and the Bill and Melinda Gates Foundation [OPP1136441].

Footnotes

Click here for additional data file.^{(1.3M, TIF)}Click here for additional data file.^{(1.2M, TIF)}Click here for additional data file.^{(995K, TIF)}Click here for additional data file.^{(20K, DOCX)}

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