Assuntos
Adenocarcinoma/secundário , Neoplasias do Ânus/patologia , Doença de Paget Extramamária/patologia , Pele/patologia , Adenocarcinoma/química , Idoso de 80 Anos ou mais , Neoplasias do Ânus/química , Biomarcadores Tumorais/análise , Biópsia , Humanos , Imuno-Histoquímica , Masculino , Invasividade Neoplásica , Doença de Paget Extramamária/química , Pele/químicaRESUMO
A method for the genetic modification of dendritic cells (DC) was previously established based on the in vitro differentiation of embryonic stem (ES) cells to DC (ES-DC). The unavailability of human ES cells genetically identical to the patients will be a problem in the future clinical application of this technology. This study attempted to establish a strategy to overcome this issue. The TAP1 or beta(2)-microglobulin (beta(2)m) gene was disrupted in 129 (H-2(b))-derived ES cells and then expression vectors for the H-2K(d) or beta(2)m-linked form of K(d) (beta2m-K(d)) were introduced, thus resulting in two types of genetically engineered ES-DC, TAP1(-/-)/K(d) ES-DC and beta(2)m(-/-)/beta(2)m-K(d) ES-DC. As intended, both of the transfectant ES-DC expressed K(d) but not the intrinsic H-2(b) haplotype-derived MHC class I. Beta(2)m(-/-)/beta(2)m-K(d) and TAP1(-/-)/K(d) ES-DC were not recognized by pre-activated H-2(b)-reactive CTL and did not prime H-2(b) reactive CTL in vitro or in vivo. Beta(2)m(-/-)/beta(2)m-K(d) ES-DC and TAP1(-/-)/K(d) ES-DC had a survival advantage in comparison to beta(2)m(+/-)/beta(2)m-K(d) ES-DC and TAP1(+/+)/K(d) ES-DC, when transferred into BALB/c mice. K(d)-restricted RSV-M2-derived peptide-loaded ES-DC could prime the epitope-specific CTL upon injection into the BALB/c mice, irrespective of the cell surface expression of intrinsic H-2(b) haplotype-encoded MHC class I. Beta(2)m(-/-)/beta(2)m-K(d) ES-DC were significantly more efficient in eliciting immunity against RSV M2 protein-expressing tumor cells than beta(2)m(+/-)/beta(2)m-K(d) ES-DC. The modification of the beta(2)m or TAP gene may therefore be an effective strategy to resolve the problem of HLA class I allele mismatch between human ES or induced pluripotent stem cells and the recipients to be treated.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Células Dendríticas/imunologia , Células-Tronco Embrionárias/imunologia , Epitopos de Linfócito T/imunologia , Antígenos H-2/genética , Ativação Linfocitária/imunologia , Linfócitos T Citotóxicos/imunologia , Microglobulina beta-2/genética , Membro 2 da Subfamília B de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/imunologia , Adenocarcinoma/prevenção & controle , Animais , Linhagem Celular , Linhagem Celular Tumoral , Células Cultivadas , Neoplasias do Colo/genética , Neoplasias do Colo/imunologia , Neoplasias do Colo/prevenção & controle , Citotoxicidade Imunológica/genética , Células Dendríticas/metabolismo , Células Dendríticas/transplante , Células-Tronco Embrionárias/metabolismo , Células-Tronco Embrionárias/transplante , Feminino , Antígenos H-2/biossíntese , Antígenos H-2/metabolismo , Haplótipos/imunologia , Teste de Histocompatibilidade , Ativação Linfocitária/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Ligação Proteica/genética , Ligação Proteica/imunologia , Linfócitos T Citotóxicos/metabolismo , Microglobulina beta-2/deficiênciaRESUMO
We report two cases of Werner's syndrome (WS). First, a 42-year-old Japanese man was referred on suspicion of systemic sclerosis (SSc) because of scleroderma-like skin atrophy and foot ulcers. Second, a 51-year-old woman with malignant fibrous histiocytoma was referred on suspicion of premature aging syndrome. Because both patients had many typical manifestations compatible with WS, we made a clinical diagnosis of WS. Genetic analyses revealed a homozygous mutation, an A deletion at nucleotide 3677 of WS gene (WRN) in the first case and a homozygous mutation, a G to C substitution at one base upstream of exon 26 of WRN in the second case. Both mutations were consistent with those previously reported in Japanese WS patients.
Assuntos
DNA Helicases/genética , Síndrome de Werner/diagnóstico , Síndrome de Werner/genética , Adulto , Primers do DNA , Diagnóstico Diferencial , Exodesoxirribonucleases , Feminino , Úlcera do Pé , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Reação em Cadeia da Polimerase , RecQ Helicases , Úlcera Cutânea , Helicase da Síndrome de WernerRESUMO
We describe a case of a 7-year-old boy diagnosed as xeroderma pigmentosum complementation group A (XPA). Severe photosensitivity developed at 5 months after birth, and at a visit to our hospital at the age of five years, multiple brownish freckles were present on the face. XPA complementing (XPAC) gene compensated the ability of DNA repair after UV-irradiation of the fibroblasts. PCR-RFLP and DNA sequencing analyses revealed compound heterozygosity for a splicing mutation (IV3 -1G => C) at the splicing acceptor site of intron 3 and a nonsense mutation (Arg228 => stop) in exon 6. The former mutation can be detected by a restriction enzyme Alw NI and the latter detected by Hph I. Neither obvious neurological symptoms nor malignant skin tumors were noted. This genotype is associated with milder clinical symptoms than homozygosity for the IV3 -1G => C mutation.