European Journal of Obstetrics & Gynecology and Reproductive Biology 182 (2014) 252–253
Contents lists available at ScienceDirect
European Journal of Obstetrics & Gynecology and Reproductive Biology journal homepage: www.elsevier.com/locate/ejogrb
Letter to the Editor—Brief Communication Preimplantation genetic diagnosis of human leukocyte antigen for X-linked immunoproliferative syndrome caused by SAP mutation
Keywords: Preimplantation genetic diagnosis (PGD) IVF X-linked immunoproliferative syndrome (XLP) SAP mutation HLA typing
Dear Editors, We would like to present a clinical case describing the generation of a non-matched human leukocyte antigen (HLA) donor in type-I of X-linked immunoproliferative syndrome (XLP), using in vitro fertilization (IVF) associated with preimplantation genetic diagnosis (PGD) for HLA typing. Hematopoietic stem cell transplantation (HSCT) is a recurrent method used for disease treatment. High polymorphism of HLA decreases the chances to identify compatible antigens. However, generation of an adequate donor can be facilitated with the utilization of PGD [1]. Association of IVF with PGD for HLA typing is a well-established procedure to conceive a child who may donate matched HSCT to its sick sibling. Generally, XLP is associated with mutated SAP gene [2], which is required for adequate function and growth of specific immune cells. In this report, we describe for the first time the molecular analysis of a family whose son was affected with XLP and PGD with HLA typing for HSCT. The mutation – c.2T > A, p/Met1Lys in SAP gene (SH2D1A) exon 1 was found on chromosome X. After DNA isolation from blood samples, 13 short tandem repeats (STRs) were selected for validation. Following the STR markers for HLA and SAP haplotype
Table 1 Primer sequences for all chosen STR markers located near the locus of SAP gene. STRs for SAP RH93240 DXS7859 DXS8096 DXS1192
F: 5-TTCATGTGATGCTTTTACCTCA-3 R: 5-TACTGGCAAGACCTGATTTCTG-3 F:5-TGCCAAGCCTCCTTTACTG-3 R: 5-ATGGAAATGGTGGGGAAAC-3 F: 5-TCACATCCAGAGAAACAGAAC-3 R: 5-GTCATGTGAGCCAGTTCTTG-3 F: 5-CAACTGCTGGAACGTTTAAA-3 R: 5-CAAAAGATTGGCTACCACTG-3
http://dx.doi.org/10.1016/j.ejogrb.2014.09.034 0301-2115/ß 2014 Published by Elsevier Ireland Ltd.
analysis, 11 markers for HLA typing (Table 1) were selected. Four additional STRs close to the mutation were chosen. Primer sequences for STR markers are provided in Table 1. Three consecutive IVF programs were done, according to standard protocols [3]. Blastomer biopsies were performed on day three. Laser technology (SaturnRI) was used to create an opening in zonas pellucidas and only one blastomer was aspirated from each 6–8 cell embryo. A total of 48 biopses from 46 embryos were analyzed, showing 14 unaffected XY and 15 carriers XX non-HLA-matched. All of them developed to blastocysts and were vitrified. In the first two cycles, frozen embryos were transferred and resulted in unsuccessful pregnancies (hCG < 1 IU). Conversely, in the third cycle, a fresh HLA-matched embryo was transferred after ensuring appropriate conditions for pregnancy establishment [3]. A single pregnancy was confirmed after the 3rd in vitro cycle, (hCG = 2960 IU). Hematopoietic stem cells 100% viable were obtained from umbilical cord. The CD34+ stem cell number in the cord blood was 5 106. The HLA-matching and SAP mutation free was confirmed after delivery of a healthy boy. The limited number of successful cases using the present preimplantation procedures highlights the importance of this report. Moreover, new sequencing techniques based on post light next generation sequencing (PLNGS) equipment totally changes the capacity, reliability and timing of analyses. In conclusion, PGD diagnostic was substituted by PLNGS technology, allowing for the generation of HLA-matched donor. References [1] Harton GL, De Rycke M, Fiorentino F, et al. ESHRE PGD consortium best practice guidelines for amplification-based PGD. Hum Reprod 2011;26:33–40. [2] Sayos J, Wu C, Morra M, et al. The X-linked lymphoproliferativedisease gene product SAP regulates signals induced through the coreceptor SLAM. Nature 1998;395:462–9. [3] Lukaszuk K, Liss J, Lukaszuk M, Maj B. Optimization of estradiol supplementation during the luteal phase improves the pregnancy rate in women undergoing in vitro fertilization-embryo transfer cycles. Fertil Steril 2005; 83(5): 1372–6.
Krzysztof Lukaszuk INVICTA Fertility and Reproductive Center, Gdansk, Poland Department of Nursing, Medical University, Gdansk, Poland Department of Gynecology and Obstetrics, Warmia and Masuria University, Olsztyn, Poland Krzysztof Kalwak Faculty of Medicine, Department of Pediatric Hematology/Oncology and Bone Marrow Transplantation, Wroclaw Medical University, Poland Sebastian Pukszta Joanna Liss INVICTA Fertility and Reproductive Center, Gdansk, Poland
Letter to the Editor—Brief Communication / European Journal of Obstetrics & Gynecology and Reproductive Biology 182 (2014) 252–253
Grzegorz Jakiel Department of Obstetrics and Gynaecology, The Medical Center of Postgraduate Education, Warsaw, Poland Izabela Woclawek-Potocka Antonio Galvao Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland
253
Tomasz Wasniewski Department of Gynecology and Obstetrics, Warmia and Masuria University, Olsztyn, Poland
E-mail addresses: [email protected], [email protected] (K. Lukaszuk). Received 18 May 2014
Contents lists available at ScienceDirect
European Journal of Obstetrics & Gynecology and Reproductive Biology journal homepage: www.elsevier.com/locate/ejogrb
Letter to the Editor—Brief Communication Preimplantation genetic diagnosis of human leukocyte antigen for X-linked immunoproliferative syndrome caused by SAP mutation
Keywords: Preimplantation genetic diagnosis (PGD) IVF X-linked immunoproliferative syndrome (XLP) SAP mutation HLA typing
Dear Editors, We would like to present a clinical case describing the generation of a non-matched human leukocyte antigen (HLA) donor in type-I of X-linked immunoproliferative syndrome (XLP), using in vitro fertilization (IVF) associated with preimplantation genetic diagnosis (PGD) for HLA typing. Hematopoietic stem cell transplantation (HSCT) is a recurrent method used for disease treatment. High polymorphism of HLA decreases the chances to identify compatible antigens. However, generation of an adequate donor can be facilitated with the utilization of PGD [1]. Association of IVF with PGD for HLA typing is a well-established procedure to conceive a child who may donate matched HSCT to its sick sibling. Generally, XLP is associated with mutated SAP gene [2], which is required for adequate function and growth of specific immune cells. In this report, we describe for the first time the molecular analysis of a family whose son was affected with XLP and PGD with HLA typing for HSCT. The mutation – c.2T > A, p/Met1Lys in SAP gene (SH2D1A) exon 1 was found on chromosome X. After DNA isolation from blood samples, 13 short tandem repeats (STRs) were selected for validation. Following the STR markers for HLA and SAP haplotype
Table 1 Primer sequences for all chosen STR markers located near the locus of SAP gene. STRs for SAP RH93240 DXS7859 DXS8096 DXS1192
F: 5-TTCATGTGATGCTTTTACCTCA-3 R: 5-TACTGGCAAGACCTGATTTCTG-3 F:5-TGCCAAGCCTCCTTTACTG-3 R: 5-ATGGAAATGGTGGGGAAAC-3 F: 5-TCACATCCAGAGAAACAGAAC-3 R: 5-GTCATGTGAGCCAGTTCTTG-3 F: 5-CAACTGCTGGAACGTTTAAA-3 R: 5-CAAAAGATTGGCTACCACTG-3
http://dx.doi.org/10.1016/j.ejogrb.2014.09.034 0301-2115/ß 2014 Published by Elsevier Ireland Ltd.
analysis, 11 markers for HLA typing (Table 1) were selected. Four additional STRs close to the mutation were chosen. Primer sequences for STR markers are provided in Table 1. Three consecutive IVF programs were done, according to standard protocols [3]. Blastomer biopsies were performed on day three. Laser technology (SaturnRI) was used to create an opening in zonas pellucidas and only one blastomer was aspirated from each 6–8 cell embryo. A total of 48 biopses from 46 embryos were analyzed, showing 14 unaffected XY and 15 carriers XX non-HLA-matched. All of them developed to blastocysts and were vitrified. In the first two cycles, frozen embryos were transferred and resulted in unsuccessful pregnancies (hCG < 1 IU). Conversely, in the third cycle, a fresh HLA-matched embryo was transferred after ensuring appropriate conditions for pregnancy establishment [3]. A single pregnancy was confirmed after the 3rd in vitro cycle, (hCG = 2960 IU). Hematopoietic stem cells 100% viable were obtained from umbilical cord. The CD34+ stem cell number in the cord blood was 5 106. The HLA-matching and SAP mutation free was confirmed after delivery of a healthy boy. The limited number of successful cases using the present preimplantation procedures highlights the importance of this report. Moreover, new sequencing techniques based on post light next generation sequencing (PLNGS) equipment totally changes the capacity, reliability and timing of analyses. In conclusion, PGD diagnostic was substituted by PLNGS technology, allowing for the generation of HLA-matched donor. References [1] Harton GL, De Rycke M, Fiorentino F, et al. ESHRE PGD consortium best practice guidelines for amplification-based PGD. Hum Reprod 2011;26:33–40. [2] Sayos J, Wu C, Morra M, et al. The X-linked lymphoproliferativedisease gene product SAP regulates signals induced through the coreceptor SLAM. Nature 1998;395:462–9. [3] Lukaszuk K, Liss J, Lukaszuk M, Maj B. Optimization of estradiol supplementation during the luteal phase improves the pregnancy rate in women undergoing in vitro fertilization-embryo transfer cycles. Fertil Steril 2005; 83(5): 1372–6.
Krzysztof Lukaszuk INVICTA Fertility and Reproductive Center, Gdansk, Poland Department of Nursing, Medical University, Gdansk, Poland Department of Gynecology and Obstetrics, Warmia and Masuria University, Olsztyn, Poland Krzysztof Kalwak Faculty of Medicine, Department of Pediatric Hematology/Oncology and Bone Marrow Transplantation, Wroclaw Medical University, Poland Sebastian Pukszta Joanna Liss INVICTA Fertility and Reproductive Center, Gdansk, Poland
Letter to the Editor—Brief Communication / European Journal of Obstetrics & Gynecology and Reproductive Biology 182 (2014) 252–253
Grzegorz Jakiel Department of Obstetrics and Gynaecology, The Medical Center of Postgraduate Education, Warsaw, Poland Izabela Woclawek-Potocka Antonio Galvao Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland
253
Tomasz Wasniewski Department of Gynecology and Obstetrics, Warmia and Masuria University, Olsztyn, Poland
E-mail addresses: [email protected], [email protected] (K. Lukaszuk). Received 18 May 2014
