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Qualitative Immunoglobulin Deficiency Causes Bacterial Infections in Patients with STAT1 Gain-of-Function Mutations

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Abstract

Purposes

STAT1 is a transduction and transcriptional regulator that functions within the classical JAK/STAT pathway. In addition to chronic mucocutaneous candidiasis, bacterial infections are a common occurrence in patients with STAT1 gain-of-function (GOF) mutations. These patients often exhibit skewing of B cell subsets; however, the impact of STAT1-GOF mutations on B cell-mediated humoral immunity remains largely unexplored. It is also unclear whether these patients with IgG within normal range require regular intravenous immunoglobulin (IVIG) therapy.

Methods

Eleven patients (harboring nine different STAT1-GOF mutations) were enrolled. Reporter assays and immunoblot analyses were performed to confirm STAT1 mutations. Flow cytometry, deep sequencing, ELISA, and ELISpot were conducted to assess the impact of STAT1-GOF on humoral immunity.

Results

All patients exhibited increased levels of phospho-STAT1 and total STAT1 protein, with two patients carrying novel mutations. In vitro assays showed that these two novel mutations were GOF mutations. Three patients with normal total IgG levels received regular IVIG infusions, resulting in effective control of bacterial infections. Four cases showed impaired affinity and specificity of pertussis toxin-specific antibodies, accompanied by reduced generation of class-switched memory B cells. Patients also had a disrupted immunoglobulin heavy chain (IGH) repertoire, coupled with a marked reduction in the somatic hypermutation frequency of switched Ig transcripts.

Conclusion

STAT1-GOF mutations disrupt B cell compartments and skew IGH characteristics, resulting in impaired affinity and antigen-specificity of antibodies and recurrent bacterial infections. Regular IVIG therapy can control these infections in patients, even those with normal total IgG levels.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

STAT1:

Signal Transducer and Activator of Transcription 1

AD:

Autosomal Dominant

GOF:

Gain-of-Function

LOF:

Loss-of-Function

IVIG:

Intravenous Immunoglobulin

IGH:

Immunoglobulin Heavy Chain

BCR:

B cell Receptor

GC:

Germinal Center

MBC:

Memory B Cell

CSR:

Class Switch Recombination

SHM:

Somatic Hypermutation

PBMC:

Peripheral Blood Mononuclear Cell

IFN:

Interferon

ELISpot:

Enzyme-linked Immunospot Assay

PT:

Pertussis Toxin

ELISA:

Enzyme-linked Immunosorbent Assay

WT:

Wild type

CADD:

Combined Annotation-Dependent Depletion

MSC:

Mutation Significance Cutoff

ASC:

Antibody-Secreting B Cell

sMBC:

Class-Switched Memory B Cell

CDR3:

Complementarity Determining Region 3

CF100:

Cumulative Frequencies of the Top 100 Abundant Clones

EBV:

Epstein-Barr Virus

CMV:

Cytomegalovirus

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Acknowledgements

The authors thank all the patients, their families, and healthy volunteers for their participation in the study.

Funding

This work was supported by the National Natural Science Foundation of China (Grant number 82070135 and 82071845), National Key R&D Program of China (Grant number 2021YFC2700804).

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Author notes

  1. Ran Chen and Huilin Mu contributed equally to this work.

Authors and Affiliations

  1. National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Child Rare Diseases in Infection and Immunity, Children’s Hospital of Chongqing Medical University, Chongqing, China

    Ran Chen, Huilin Mu, Lina Zhou, Zhiyong Zhang, Xuemei Tang, Yanjun Jia, Xiaodong Zhao & Yunfei An

  2. Department of Rheumatology and Immunology, Children’s Hospital of Chongqing Medical University, No. 136, Zhongshan 2nd Road, Yuzhong District, Chongqing, 400014, China

    Zhiyong Zhang, Xuemei Tang, Xiaodong Zhao & Yunfei An

  3. Department of Rheumatology and Immunology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Xuemei Chen

  4. Department of Pediatrics, Graduate School of Biomedical and Health Sciences, 1-2-3 Kasumi, Minami-Ku, Hiroshima, 734-8551, Japan

    Miyuki Tsumura & Satoshi Okada

  5. Department of Nephrology and Immunology, Guiyang Maternal & Child Health Care Hospital, Guiyang, China

    Xinhui Jiang

  6. Institute of Immunology, PLA, Third Military Medical University, Chongqing, 400038, People’s Republic of China

    Yongwen Chen

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  1. Ran Chen
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Contributions

Ran Chen and Huilin Mu mainly performed the study and wrote the manuscript. Xuemei Chen and Miyuki Tsumura performed some experiments. Lina Zhou and Xinhui Jiang revised the manuscript. Zhiyong Zhang, Xuemei Tang, Yongwen Chen and Yanjun Jia collected the samples and characterized patient clinical information. Satoshi Okada revised the manuscript, and provided advice on the design of the study. Xiaodong Zhao and Yunfei An designed the research. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Satoshi Okada, Xiaodong Zhao or Yunfei An.

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Ethics Approval

The study was performed following Declaration of Helsinki and approved by the Institutional Review Board of Children’s Hospital of Chongqing Medical University (2021–138).

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Written informed consents for involvement in this study were provided by all enrolled subjects’ parents.

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Written informed consent for publication of the study was obtained from all enrolled subjects’ parents.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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Chen, R., Mu, H., Chen, X. et al. Qualitative Immunoglobulin Deficiency Causes Bacterial Infections in Patients with STAT1 Gain-of-Function Mutations. J Clin Immunol 44, 124 (2024). https://doi.org/10.1007/s10875-024-01720-x

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