1. Follow-up of a patient with CERNUNNOS deficiency Edyta Heropolitańska-Pliszka Immunology Department Children’s Memorial Health Institute Warsaw, Poland Children’s Memorial Health Institute Warsaw, Poland

2. CERNUNNOS / XLF a new type of radiosensitive a new type of radiosensitive T-B-NK+ SCID T-B-NK+ SCID cernu nnos

3. Cernunnos deficiency core components consanguinity consanguinity recurrent bacterial and opportunistic infections recurrent bacterial and opportunistic infections microcephaly microcephaly severe growth retardation severe growth retardation dysmorfic features dysmorfic features hypogammaglobulinemia IgG and IgA hypogammaglobulinemia IgG and IgA fluctuating level of IgM fluctuating level of IgM mild to severe B and T lymphopenia, NK cells not affected mild to severe B and T lymphopenia, NK cells not affected all T cells of memory phenotype (CD45 RO+) all T cells of memory phenotype (CD45 RO+) impaired response of T cells to PHA impaired response of T cells to PHA chromosomal translocations in lymphocytes chromosomal translocations in lymphocytes increased radiosensitivity increased radiosensitivity

4. 8-year-old boy irrelevant family history irrelevant family history recurrent bacterial and viral infections of upper and lower respiratory tract since infancy period (otitis, bronchitis and pneumonitis every 2 months) recurrent bacterial and viral infections of upper and lower respiratory tract since infancy period (otitis, bronchitis and pneumonitis every 2 months) significant microcephaly (-5 SD) significant microcephaly (-5 SD) birdlike face birdlike face growth retardation (<3 pc) growth retardation (<3 pc)

5. Immunologic investigation IgG 120-530 mg/dl, IgG 120-530 mg/dl, IgA 6-18 mg/dl, IgA 6-18 mg/dl, IgM 921-1390 mg/dl IgM 921-1390 mg/dl CD19+ 4,3% - 81 cells/ul CD19+ 4,3% - 81 cells/ul CD3+ 32% - 380 cells/ul CD3+ 32% - 380 cells/ul CD16+56+ 43% - 665 cells/ul CD16+56+ 43% - 665 cells/ul CD4+ CD45RO+ 91,8% - 600 cells/ul CD4+ CD45RO+ 91,8% - 600 cells/ul CD8+ CD45RO+ 64.7% - 466 cells/ul CD8+ CD45RO+ 64.7% - 466 cells/ul PHA 2635+/-7 PHA 2635+/-7 anty-CD3 838+/-65 anty-CD3 838+/-65 spontaneous chromosomal fragility spontaneous chromosomal fragility increased radiosensitivity of patient’s fibroblasts increased radiosensitivity of patient’s fibroblasts

6. CERNUNNOS deficiency homozygous stop codon mutation Y167X

7. HSCT March 2006 - MUD PBPC (CD34+ 12,7x10 6 /kg b.m) March 2006 - MUD PBPC (CD34+ 12,7x10 6 /kg b.m) Conditioning regimen reduced because of breakage of chromosomes (FluCyATG) Conditioning regimen reduced because of breakage of chromosomes (FluCyATG) Early outcome: Early outcome: +12 day - GVHD II stage in skin (steroids) +12 day - GVHD II stage in skin (steroids) +17 day - good haematological reconstitution +17 day - good haematological reconstitution +27 day - complete chimerism +27 day - complete chimerism

8. + 62 day good clinical condition, no symptoms of infection good clinical condition, no symptoms of infection complete chimerism, no signs of GVHD complete chimerism, no signs of GVHD immunosupression: CsA and steroids (Encorton 0,5 mg/kg b.m.) immunosupression: CsA and steroids (Encorton 0,5 mg/kg b.m.) prophylaxis: Co- trimoxazole, Azithromycin, Acyclovir, Fluconazole prophylaxis: Co- trimoxazole, Azithromycin, Acyclovir, Fluconazole

9. +155 day hemorrhagic cystitis of unknown origin (polioma-, adeno-, CMV, EBV excluded) hemorrhagic cystitis of unknown origin (polioma-, adeno-, CMV, EBV excluded) treatment: forced diuresis, IVIG substitution, intravenous Acyclovir treatment: forced diuresis, IVIG substitution, intravenous Acyclovir +210 day – good clinical condition, normal blood and urine tests, complete chimerism +210 day – good clinical condition, normal blood and urine tests, complete chimerism

10. +385 day Protracted fever with lymphadenopathy, skin changes (chronic GVHD) Protracted fever with lymphadenopathy, skin changes (chronic GVHD) CMV PCR (-) CMV PCR (-) Aspergillus ELISA 0,452 PCR (-) Aspergillus ELISA 0,452 PCR (-) Cryptosporidium sp. PCR (-) Cryptosporidium sp. PCR (-) EBV DNA 140-560 copies EBV DNA 140-560 copies Diagnosis: EBV infection Diagnosis: EBV infection Treatment: Azithromycin, Co- trimoxazol, Acyclovir, Worikonazol, IVIG every 10 days, UDCA, Gancyclovir Treatment: Azithromycin, Co- trimoxazol, Acyclovir, Worikonazol, IVIG every 10 days, UDCA, Gancyclovir WBC – 7,8 K/ul WBC – 7,8 K/ul N-29% L-35% N-29% L-35% E-27% AL-2% E-27% AL-2% M-7% M-7% Hgb – 10,2 g/dl Hgb – 10,2 g/dl Plt - 135 K/ul Plt - 135 K/ul GOT – 110 U/l GOT – 110 U/l GGTP – 335 U/l GGTP – 335 U/l CRP - 1,2 mg/dl CRP - 1,2 mg/dl LDH – 245 U/l LDH – 245 U/l

11. +425 day - last follow-up IgG 590 mg/dl, IgG 590 mg/dl, IgA 41 mg/dl, IgA 41 mg/dl, IgM 60 mg/dl IgM 60 mg/dl CD19+ 12,3% - 405 cells/ul CD19+ 12,3% - 405 cells/ul CD3+ 69% - 2310 cells/ul CD3+ 69% - 2310 cells/ul CD16+56+ 13% - 469 cells/ul CD16+56+ 13% - 469 cells/ul Alone 337+/-80 Alone 337+/-80 PHA 16871+/-645 PHA 16871+/-645 CD3 16199+/-125 CD3 16199+/-125 Complete chimerism Complete chimerism

12. Thank you for attention