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Human IgG1 (B-cell receptor) Fc peptide glycan analysis, LC-MS

ABSTRACT: Since few data are available on glycans expressed by human BCRs, we aimed to analyze the Fc glycans of membrane-bound IgG after BCR capture and tryptic digestion by LC-MS. For this purpose, human B-cell lines from Burkitt Lymphoma (Ramos) expressing IgG1-BCRs in the presence or absence of Fc glycans (FcG) were analyzed. The B-cell lines were generated by transduction of the BCR-negative MDL-AID KO cell line with N(297) or mutant Q(297) IgG-BCRs. The BCR sequences used were derived from single-cell sorted human B cells isolated from patients with the autoimmune disease rheumatoid arthritis. Two BCRs were directed towards citrullinated antigens (2G9 and 3F3), while a third BCR was directed against tetanus toxoid (D2). In addition, we analyzed the Fc glycan profile of IgG secreted (sIgG) by human Ramos B cells for comparison.

INSTRUMENT(S): maXis

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): B Cell, Cell Culture

DISEASE(S): Rheumatoid Arthritis

SUBMITTER: Theresa Kissel

LAB HEAD: Rene E.M. Toes

PROVIDER: PXD047089 | Pride | 2023-12-11

REPOSITORIES: Pride

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			Action	DRS
	2021GLY00003MX5_3F3_FcG_negative.mzXML	Mzxml
	2021GLY00003MX5_3F3_FcG_positive.mzXML	Mzxml
	2021GLY00003MX5_MDL_AID_KO.mzXML	Mzxml
	2022GLY00056MX5_D2_FcG_negative.mzXML	Mzxml
	2022GLY00056MX5_D2_FcG_positive.mzXML	Mzxml

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Project description:The transplacental transfer of maternal IgG to the developing fetus is critical for infant protection against infectious pathogens in the first year of life. However, factors that modulate the transplacental transfer efficiency of maternal IgG that could be harnessed for maternal vaccine design remain largely undefined. HIV-infected women have impaired placental IgG transfer, yet the mechanism underlying this impaired transfer is unknown, presenting an opportunity to explore factors that contribute to the efficiency of placental IgG transfer. We measured the transplacental transfer efficiency of maternal HIV and other pathogen-specific IgG in historical U.S. (n=120) and Malawian (n=47) cohorts of HIV-infected mothers and their HIV58 exposed uninfected and HIV-infected infants. We then examined the role of maternal HIV disease progression, infant factors, placental Fc receptor expression, and IgG Fc region subclass and glycan signatures and their association with transplacental transfer efficiency of maternal antigen-specific IgG. We established 3 distinct phenotypes of placental IgG transfer efficiency in HIV-infected women, including: 1) efficient transfer of the majority of antigen-specific IgG populations; 2) generally poor IgG transfer phenotype that was strongly associated with maternal CD4+ T cell counts, hypergammaglobulinemia, and frequently yielded non-protective levels of vaccine-specific IgG; and 3) variable transfer of IgG across distinct antigen specificities. Interestingly, maternal IgG characteristics, such as binding to placentally expressed Fc receptors FcgRIIa and FcgRIIIa, IgG subclass frequency, and Fc region glycan profiles were associated with placental IgG transfer efficiency. These maternal IgG transplacental transfer determinants were distinct among different antigen-specific IgG populations. Our findings suggest that in HIV-infected women, both maternal disease progression and Fc region characteristics modulate the selective placental transfer of distinct IgG subpopulations, with implications for both the health of HIV-exposed uninfected infants and maternal vaccine design.

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Human IgG1 (B-cell receptor) Fc peptide glycan analysis, LC-MS

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