Abstract
Zeta-chain associated protein kinase 70 kDa (ZAP70) combined immunodeficiency (CID) is an autosomal recessive severe immunodeficiency that is characterized by abnormal T-cell receptor signaling. Children with the disorder typically present during the first year of life with diarrhea, failure to thrive, and recurrent bacterial, viral, or opportunistic infections. To date, the only potential cure is hematopoietic stem cell transplant (HSCT). The majority of described mutations causing disease occur in the homozygous state, though heterozygotes are reported without a clear understanding as to how the individual mutations interact to cause disease. This case describes an infant with novel ZAP-70 deficiency mutations involving the SH2 and kinase domains cured with allogeneic HSCT utilizing a reduced-intensity conditioning regimen and graft manipulation. We then were able to further elucidate the molecular signaling alterations imparted by these mutations that lead to altered immune function. In order to examine the effect of these novel compound ZAP70 heterozygous mutations on T cells, Jurkat CD4+ T cells were transfected with either wild type, or with individual ZAP70 R37G and A507T mutant constructs. Downstream TCR signaling events and protein localization results link these novel mutations to the expected immunological outcome as seen in the patient’s primary cells. This study further characterizes mutations in the ZAP70 gene as combined immunodeficiency and the clinical phenotype.
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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.
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Acknowledgements
We are grateful for the patient, her family, and the healthy donors that contributed to this study. We would like to thank Emilia Scharrig, Margaret McCormick, and Waldo Spessott for their technical assistance and training.
Funding
This work was supported by the R01 AI123538 grant to C.G.G. and T32 AI 134646–4 grant to N.B., and Pediatric Award in Clinical Research (PACR) Pilot funds from Thomas Jefferson University and Nemours Children’s Hospital to C.G.G. and J.C.W. We would like to thank the Bioimaging Shared Resource of the Sidney Kimmel Cancer Center (NCI 5 P30 CA-56036).
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N.B., J.C.W., and C.G.G. designed the study. N.B., J.C.W., and M.L.S. performed the experiments and collected the data. N.B. and C.G.G. analyzed and interpreted the data. J.C.W., M.T., T.L., and E.C. carried out the clinical diagnosis and management. N.B., J.C.W., and C.G.G. wrote the manuscript and all authors contributed to editing.
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This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the institution review boards of Nemours (IRB# 349465).
and Thomas Jefferson University (IRB # 18F.708) Dated 05/26/2021.
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Written consent was obtained from either the parents or from all individual participants included in the study.
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10875_2023_1608_MOESM1_ESM.pdf
Supplementary file1 (PDF 936 KB) Analysis of T-Lymphocyte Mitogen Stimulation using the clinical tests pokeweed (PWM) and phytohemagglutinin (PHA). Pre- and Post-transplant patient PBMCs were sent out for evaluation for their response to either Media alone, PWM or PHA using a CLIA-certified test at the Mayo Clinic. Plots show that both pre-transplant CD45+ cells and CD3+ patient cells were unable to proliferate upon incubation with PWM or PHA. Interestingly, Post-transplant cells were able to restore their proliferation activity within the normal reference range as shown in Table 1.
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Benavides, N., White, J.C., Sanmillan, M.L. et al. Novel Compound Heterozygous ZAP70 R37G A507T Mutations in Infant with Severe Immunodeficiency. J Clin Immunol 44, 27 (2024). https://doi.org/10.1007/s10875-023-01608-2
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DOI: https://doi.org/10.1007/s10875-023-01608-2