Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodies
Abstract Pre- and post-transplantation anti-MICA antibody detection development are associated with an increased rejection risk and low graft survival. We previously generated HLA class I null HEK-293T using CRISPR/Cas9, while MICA and MICB genes were removed in this study. A panel of 11 cell lines...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2021-08-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-95058-8 |
| _version_ | 1818696306889064448 |
|---|---|
| author | Ji-Ho Jeon In-Cheol Baek Cheol-Hwa Hong Ki Hyun Park Hyeyoung Lee Eun-Jee Oh Tai-Gyu Kim |
| author_facet | Ji-Ho Jeon In-Cheol Baek Cheol-Hwa Hong Ki Hyun Park Hyeyoung Lee Eun-Jee Oh Tai-Gyu Kim |
| author_sort | Ji-Ho Jeon |
| collection | DOAJ |
| description | Abstract Pre- and post-transplantation anti-MICA antibody detection development are associated with an increased rejection risk and low graft survival. We previously generated HLA class I null HEK-293T using CRISPR/Cas9, while MICA and MICB genes were removed in this study. A panel of 11 cell lines expressing single MICA alleles was established. Anti-MICA antibody in the sera of kidney transplant patients was determined using flow cytometric method (FCM) and the Luminex method. In the 44 positive sera, the maximum FCM value was 2879 MFI compared to 28,135 MFI of Luminex method. Eleven sera (25%) were determined as positive by FCM and 32 sera (72%) were positive by the Luminex method. The sum of total MICA antigens, MICA*002, *004, *009, *019, and *027 correlation showed a statistically significant between the two methods (P = 0.0412, P = 0.0476, P = 0.0019, P = 0.0098, P = 0.0467, and P = 0.0049). These results demonstrated that HEK-293T-based engineered cell lines expressing single MICA alleles were suitable for measuring specific antibodies against MICA antigens in the sera of transplant patients. Studies of antibodies to MICA antigens may help to understand responses in vivo and increase clinical relevance at the cellular level such as complement-dependent cytotoxicity. |
| first_indexed | 2024-12-17T13:59:17Z |
| format | Article |
| id | doaj.art-c13fc26bf6434a42ab6e2b7ee7419048 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-17T13:59:17Z |
| publishDate | 2021-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-c13fc26bf6434a42ab6e2b7ee74190482022-12-21T21:45:50ZengNature PortfolioScientific Reports2045-23222021-08-0111111210.1038/s41598-021-95058-8Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodiesJi-Ho Jeon0In-Cheol Baek1Cheol-Hwa Hong2Ki Hyun Park3Hyeyoung Lee4Eun-Jee Oh5Tai-Gyu Kim6Department of Microbiology, College of Medicine, The Catholic University of KoreaHematopoietic Stem Cell Bank, College of Medicine, The Catholic University of KoreaDepartment of Microbiology, College of Medicine, The Catholic University of KoreaDepartments of Laboratory Medicine, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of KoreaDepartment of Laboratory Medicine, International St. Mary’s Hospital, College of Medicine, Catholic Kwandong UniversityDepartments of Laboratory Medicine, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of KoreaDepartment of Microbiology, College of Medicine, The Catholic University of KoreaAbstract Pre- and post-transplantation anti-MICA antibody detection development are associated with an increased rejection risk and low graft survival. We previously generated HLA class I null HEK-293T using CRISPR/Cas9, while MICA and MICB genes were removed in this study. A panel of 11 cell lines expressing single MICA alleles was established. Anti-MICA antibody in the sera of kidney transplant patients was determined using flow cytometric method (FCM) and the Luminex method. In the 44 positive sera, the maximum FCM value was 2879 MFI compared to 28,135 MFI of Luminex method. Eleven sera (25%) were determined as positive by FCM and 32 sera (72%) were positive by the Luminex method. The sum of total MICA antigens, MICA*002, *004, *009, *019, and *027 correlation showed a statistically significant between the two methods (P = 0.0412, P = 0.0476, P = 0.0019, P = 0.0098, P = 0.0467, and P = 0.0049). These results demonstrated that HEK-293T-based engineered cell lines expressing single MICA alleles were suitable for measuring specific antibodies against MICA antigens in the sera of transplant patients. Studies of antibodies to MICA antigens may help to understand responses in vivo and increase clinical relevance at the cellular level such as complement-dependent cytotoxicity.https://doi.org/10.1038/s41598-021-95058-8 |
| spellingShingle | Ji-Ho Jeon In-Cheol Baek Cheol-Hwa Hong Ki Hyun Park Hyeyoung Lee Eun-Jee Oh Tai-Gyu Kim Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodies Scientific Reports |
| title | Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodies |
| title_full | Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodies |
| title_fullStr | Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodies |
| title_full_unstemmed | Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodies |
| title_short | Establishment of HLA class I and MICA/B null HEK-293T panel expressing single MICA alleles to detect anti-MICA antibodies |
| title_sort | establishment of hla class i and mica b null hek 293t panel expressing single mica alleles to detect anti mica antibodies |
| url | https://doi.org/10.1038/s41598-021-95058-8 |
| work_keys_str_mv | AT jihojeon establishmentofhlaclassiandmicabnullhek293tpanelexpressingsinglemicaallelestodetectantimicaantibodies AT incheolbaek establishmentofhlaclassiandmicabnullhek293tpanelexpressingsinglemicaallelestodetectantimicaantibodies AT cheolhwahong establishmentofhlaclassiandmicabnullhek293tpanelexpressingsinglemicaallelestodetectantimicaantibodies AT kihyunpark establishmentofhlaclassiandmicabnullhek293tpanelexpressingsinglemicaallelestodetectantimicaantibodies AT hyeyounglee establishmentofhlaclassiandmicabnullhek293tpanelexpressingsinglemicaallelestodetectantimicaantibodies AT eunjeeoh establishmentofhlaclassiandmicabnullhek293tpanelexpressingsinglemicaallelestodetectantimicaantibodies AT taigyukim establishmentofhlaclassiandmicabnullhek293tpanelexpressingsinglemicaallelestodetectantimicaantibodies |