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Combined immunodeficiency due to MALT1 mutations, treated by hematopoietic cell transplantation.

Punwani D, Wang H, Chan AY, Cowan MJ, Mallott J, Sunderam U, Mollenauer M, Srinivasan R, Brenner SE, Mulder A, Claas FH, Weiss A, Puck JM - J. Clin. Immunol. (2015)

Bottom Line: No defects were found in multiple genes associated with severe combined immunodeficiency.Our nonconsanguineous patient with early onset profound combined immunodeficiency and immune dysregulation due to compound heterozygous MALT1 mutations extends the clinical and immunologic phenotype reported in 2 prior families.Clinical cure was achieved with mixed chimerism after nonmyeloablative conditioning and HCT.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, University of California San Francisco School of Medicine, and UCSF Benioff Children's Hospital, Box 0519, 513 Parnassus Avenue, HSE-301A, San Francisco, CA, 94143-0519, USA.

ABSTRACT

Purpose: A male infant developed generalized rash, intestinal inflammation and severe infections including persistent cytomegalovirus. Family history was negative, T cell receptor excision circles were normal, and engraftment of maternal cells was absent. No defects were found in multiple genes associated with severe combined immunodeficiency. A 9/10 HLA matched unrelated hematopoietic cell transplant (HCT) led to mixed chimerism with clinical resolution. We sought an underlying cause for this patient's immune deficiency and dysregulation.

Methods: Clinical and laboratory features were reviewed. Whole exome sequencing and analysis of genomic DNA from the patient, parents and 2 unaffected siblings was performed, revealing 2 MALT1 variants. With a host-specific HLA-C antibody, we assessed MALT1 expression and function in the patient's post-HCT autologous and donor lymphocytes. Wild type MALT1 cDNA was added to transformed autologous patient B cells to assess functional correction.

Results: The patient had compound heterozygous DNA variants affecting exon 10 of MALT1 (isoform a, NM_006785.3), a maternally inherited splice acceptor c.1019-2A > G, and a de novo deletion of c.1059C leading to a frameshift and premature termination. Autologous lymphocytes failed to express MALT1 and lacked NF-κB signaling dependent upon the CARMA1, BCL-10 and MALT1 signalosome. Transduction with wild type MALT1 cDNA corrected the observed defects.

Conclusions: Our nonconsanguineous patient with early onset profound combined immunodeficiency and immune dysregulation due to compound heterozygous MALT1 mutations extends the clinical and immunologic phenotype reported in 2 prior families. Clinical cure was achieved with mixed chimerism after nonmyeloablative conditioning and HCT.

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a, Intracellular phospho-NF-κB and IκB in gated unstimulated (gray shading) vs. PMA and ionomycin stimulated (black line) naïve and memory CD3+ T cells and CD19+ B cells from the patient, including patient autologous (TRA2G9+) and donor-derived (TRA2G9-) cells; also analyzed were cells from the mother and a healthy control. b, T cell expression of intracellular IL-2 (y-axis) and IFN-γ (x-axis) without (left panels) and with (right panels) PMA plus ionomycin stimulation. c, Analysis of NF-κB phosphorylation and IκB degradation without (gray shading) or with (black lines) stimulation with PMA and ionomycin. Upper panels, control EBV B cells; middle panels, patient autologous EBV B cells transduced with empty MP283 lentivirus; bottom panels, patient autologous EBV B cells transduced with MP283-MALT1 lentivirus. All data representative of 3 independent experiments
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Fig3: a, Intracellular phospho-NF-κB and IκB in gated unstimulated (gray shading) vs. PMA and ionomycin stimulated (black line) naïve and memory CD3+ T cells and CD19+ B cells from the patient, including patient autologous (TRA2G9+) and donor-derived (TRA2G9-) cells; also analyzed were cells from the mother and a healthy control. b, T cell expression of intracellular IL-2 (y-axis) and IFN-γ (x-axis) without (left panels) and with (right panels) PMA plus ionomycin stimulation. c, Analysis of NF-κB phosphorylation and IκB degradation without (gray shading) or with (black lines) stimulation with PMA and ionomycin. Upper panels, control EBV B cells; middle panels, patient autologous EBV B cells transduced with empty MP283 lentivirus; bottom panels, patient autologous EBV B cells transduced with MP283-MALT1 lentivirus. All data representative of 3 independent experiments

Mentions: Phosphorylation of NF-κB and degradation of IκB following stimulation were analyzed to indicate the status of the CBM signalosome in PBMCs from the patient, his mother and a control. The patient’s autologous cells, identified by staining with the TRA2G9 antibody, were unable to phosphorylate NF-κB or degrade IκB (Fig. 3a, left and right panels, respectively), while cells from the mother and control cells demonstrated equivalent levels of NF-κB phosphorylation and IκB degradation in naïve and memory T cells as well as B cells. As noted above, too few donor-derived naïve T cells were detected in the patient’s blood to analyze. Phosphorylation of Erk, P38 and S6, each independent of the CBM signalosome, was intact in autologous patient cells, demonstrating selectivity of the MALT1 defect (Suppl Figure 1).Fig. 3


Combined immunodeficiency due to MALT1 mutations, treated by hematopoietic cell transplantation.

Punwani D, Wang H, Chan AY, Cowan MJ, Mallott J, Sunderam U, Mollenauer M, Srinivasan R, Brenner SE, Mulder A, Claas FH, Weiss A, Puck JM - J. Clin. Immunol. (2015)

a, Intracellular phospho-NF-κB and IκB in gated unstimulated (gray shading) vs. PMA and ionomycin stimulated (black line) naïve and memory CD3+ T cells and CD19+ B cells from the patient, including patient autologous (TRA2G9+) and donor-derived (TRA2G9-) cells; also analyzed were cells from the mother and a healthy control. b, T cell expression of intracellular IL-2 (y-axis) and IFN-γ (x-axis) without (left panels) and with (right panels) PMA plus ionomycin stimulation. c, Analysis of NF-κB phosphorylation and IκB degradation without (gray shading) or with (black lines) stimulation with PMA and ionomycin. Upper panels, control EBV B cells; middle panels, patient autologous EBV B cells transduced with empty MP283 lentivirus; bottom panels, patient autologous EBV B cells transduced with MP283-MALT1 lentivirus. All data representative of 3 independent experiments
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4352191&req=5

Fig3: a, Intracellular phospho-NF-κB and IκB in gated unstimulated (gray shading) vs. PMA and ionomycin stimulated (black line) naïve and memory CD3+ T cells and CD19+ B cells from the patient, including patient autologous (TRA2G9+) and donor-derived (TRA2G9-) cells; also analyzed were cells from the mother and a healthy control. b, T cell expression of intracellular IL-2 (y-axis) and IFN-γ (x-axis) without (left panels) and with (right panels) PMA plus ionomycin stimulation. c, Analysis of NF-κB phosphorylation and IκB degradation without (gray shading) or with (black lines) stimulation with PMA and ionomycin. Upper panels, control EBV B cells; middle panels, patient autologous EBV B cells transduced with empty MP283 lentivirus; bottom panels, patient autologous EBV B cells transduced with MP283-MALT1 lentivirus. All data representative of 3 independent experiments
Mentions: Phosphorylation of NF-κB and degradation of IκB following stimulation were analyzed to indicate the status of the CBM signalosome in PBMCs from the patient, his mother and a control. The patient’s autologous cells, identified by staining with the TRA2G9 antibody, were unable to phosphorylate NF-κB or degrade IκB (Fig. 3a, left and right panels, respectively), while cells from the mother and control cells demonstrated equivalent levels of NF-κB phosphorylation and IκB degradation in naïve and memory T cells as well as B cells. As noted above, too few donor-derived naïve T cells were detected in the patient’s blood to analyze. Phosphorylation of Erk, P38 and S6, each independent of the CBM signalosome, was intact in autologous patient cells, demonstrating selectivity of the MALT1 defect (Suppl Figure 1).Fig. 3

Bottom Line: No defects were found in multiple genes associated with severe combined immunodeficiency.Our nonconsanguineous patient with early onset profound combined immunodeficiency and immune dysregulation due to compound heterozygous MALT1 mutations extends the clinical and immunologic phenotype reported in 2 prior families.Clinical cure was achieved with mixed chimerism after nonmyeloablative conditioning and HCT.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, University of California San Francisco School of Medicine, and UCSF Benioff Children's Hospital, Box 0519, 513 Parnassus Avenue, HSE-301A, San Francisco, CA, 94143-0519, USA.

ABSTRACT

Purpose: A male infant developed generalized rash, intestinal inflammation and severe infections including persistent cytomegalovirus. Family history was negative, T cell receptor excision circles were normal, and engraftment of maternal cells was absent. No defects were found in multiple genes associated with severe combined immunodeficiency. A 9/10 HLA matched unrelated hematopoietic cell transplant (HCT) led to mixed chimerism with clinical resolution. We sought an underlying cause for this patient's immune deficiency and dysregulation.

Methods: Clinical and laboratory features were reviewed. Whole exome sequencing and analysis of genomic DNA from the patient, parents and 2 unaffected siblings was performed, revealing 2 MALT1 variants. With a host-specific HLA-C antibody, we assessed MALT1 expression and function in the patient's post-HCT autologous and donor lymphocytes. Wild type MALT1 cDNA was added to transformed autologous patient B cells to assess functional correction.

Results: The patient had compound heterozygous DNA variants affecting exon 10 of MALT1 (isoform a, NM_006785.3), a maternally inherited splice acceptor c.1019-2A > G, and a de novo deletion of c.1059C leading to a frameshift and premature termination. Autologous lymphocytes failed to express MALT1 and lacked NF-κB signaling dependent upon the CARMA1, BCL-10 and MALT1 signalosome. Transduction with wild type MALT1 cDNA corrected the observed defects.

Conclusions: Our nonconsanguineous patient with early onset profound combined immunodeficiency and immune dysregulation due to compound heterozygous MALT1 mutations extends the clinical and immunologic phenotype reported in 2 prior families. Clinical cure was achieved with mixed chimerism after nonmyeloablative conditioning and HCT.

Show MeSH
Related in: MedlinePlus