Extremely polymorphic genes of major histocompatibility complex (MHC) play a significant role in the function of immune system by recognizing heterogeneous particles, mainly pathogenic origin. Previous research on various vertebrate species indicates that MHC influences individual body odour and mate choice preferences. Many individuals tend to prefer MHC dissimilar partner so that warrants them an offspring resistant against wider spectrum of infections. Research on MHC-related mate preferences in humans, however, is inconclusive to date. Several studies indicate that women not taking hormonal contraceptives prefer the smell of MHC dissimilar partners while other studies have not come to this conclusion. This can be caused by the absence of potentially influencing factors like the menstrual cycle phase. The aim of this study was to test MHC-similarity mate choice preferences in odour, facial and vocal modalities. In particular, we focused on a potential effect of hormonal contraception. Furtermore, we tested preferential shifts across the menstrual cycle by comparing women's preferences in the follicular and the luteal phase in pill and non-pill users. A group of 52 women in different phases of their menstrual cycle rated odour samples, photos and vocal recordings taken from 51 men. All...

京都大学 / 0048 / 新制・論文博士 / 博士(農学) / 乙第12986号 / 論農博第2826号 / 新制||農||1038(附属図書館) / 学位論文||H28||N4961(農学部図書室) / 32456 / 名古屋大学大学院農学研究科生化学制御専攻 / (主査)教授 祝前 博明, 教授 今井 裕, 教授 廣岡 博之 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

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京都大学 / 新制・課程博士 / 博士(医学) / 甲第24991号 / 医博第5025号 / 新制||医||1069(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 生田 宏一, 教授 椛島 健治, 教授 上野 英樹 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

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Development of techniques for observing variability at the major histocompatibility complex (MHC) of fishes could prove an important first step in understanding the genetic bases of disease resistance. In this study, using goldfish (Carassius auratus) as a model system, three approaches to generating antisera to putative MHC molecules and two methods for detecting antibody reactivities were evaluated. Seven full-sib families were produced, and red blood cells (RBCs) of goldfish family members were screened for reactivity with a panel of absorbed antisera. The antisera panel consisted of fish anti-fish, chicken anti-chicken, and chicken anti-fish antisera. The fish anti-fish antisera was produced by injecting RBCs from each parent into its mate, and the chicken anti-fish antisera was produced by injecting parental goldfish RBCs into chickens. The chicken anti-chicken antisera were obtained from a genetics laboratory where MHC-specific antisera had been prepared previously. The pattern of presence or absence of agglutination upon mixing with the respective reagents in this panel of antisera was regarded as the phenotype of the individual tested. Agglutinations observed macroscopically or microscopically were easily scored as positive or negative. Particular phenotypes were observed among individuals both within and between families. The large numbers of phenotypes observed may indicate: (1) the need for additional absorptions in the preparation of antisera, or (2) segregation of additional sets of phenotypic MHC haplotypes in the tetraploid goldfish. The utility of chicken anti-chicken reagents in serotyping of fish was demonstrated. Use of the traditional approach to conducting hemagglutination assays limited the number of assays executed because of the amount of blood required. In order to minimize the sample volumes required, antibody reactivities were evaluated by flow cytometry employing appropriate fluorescein labeled antibodies. Using this approach, scoring of positive and negative results was equivocal, and results did not always agree with those scored by hemagglutination assays. Results of this study strongly suggest that the development of immune allo- and xeno-antisera and use of hemagglutination assays can be used to characterize genetic variability of the MHC of fishes. Understanding of immunogenetic variability in fishes could be used to develop strains resistant to economically important fish pathogens. / Master of Science

付記する学位プログラム名: 霊長類学・ワイルドライフサイエンス・リーディング大学院 / 京都大学 / 新制・課程博士 / 博士(理学) / 甲第25153号 / 理博第5060号 / 新制||理||1721(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 村山 美穂, 教授 平田 聡, 教授 三谷 曜子 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

My dissertation hypothesis is that bovine trophoblast cells express cell-surface and secreted non-classical major histocompatibility complex class I (MHC-Ib) proteins which inhibit NK cells and other leukocytes by binding to inhibitory receptors (e.g., LILRB1, LILRB2, KIR2DL4, and/or CD94/NKG2A). Extremely polymorphic and ubiquitously expressed classical MHC class I (MHC-Ia) proteins, which present foreign antigenic peptides to CD8+ T lymphocytes, are involved in acceptance or rejection of tissue grafts. Non-classical MHC class I (MHC-Ib) glycoproteins, such as Human Leukocyte Antigen-G (HLA-G) and murine Qa-2, are important modulators of the maternal immune system during pregnancy. MHC-Ib proteins are: (a) oligomorphic or monomorphic, (b) expressed in specific tissues under specific condtions, and (c) produced as surface and/or soluble isoforms due to alternative splicing. Third trimester-bovine trophoblast cells express both MHC-Ia and MHC-Ib proteins. The MHC-Ib proteins expressed by trophoblast cells during the third trimester of pregnancy are encoded by four bovine leukocyte antigen (BoLA) loci: BoLA-NC1, BoLA-NC2, BoLA-NC3, and BoLA-NC4. Two MHC-Ia (N*01701 and N*01802) and three MHC-Ib (NC1*00501, NC3*00101 and NC4*00201) proteins showed cell-surface expression in transfection studies performed in murine P815 and human K562 cells. Two additional isoforms, NC1*00401 and NC2*00102, were not detected on the surface of these cells. Nevertheless, both class Ia proteins, N*01701 and N*01802, and five class Ib proteins, NC1*00401, NC1*00501, NC2*00102, NC3*00101, and NC4*00201, were detected in crude cell lysates on Western blots. Precipitation of proteins from culture supernatants showed that cell-surface MHC-Ia (N*01701 and N*01802) and MHC-Ib proteins (NC1*00501, NC3*00101, and NC4*00201) are shed from the surface of these cells into the media. The mechanism of shedding of these proteins is, however, not known. Monoclonal antibodies W6/32, IL-A88, H1A, H6A, H11A, H58A, and PT-85A recognized surface MHC-I isoforms with varying affinity. We were able to develop a sandwich enzyme-linked immunosorbent assay (ELISA) using either H1A or IL-A88 antibody as the capture antibody and the W6/32 antibody for detection. We produced monoclonal antibodies against cattle NC1*00501 and NC3*00101 proteins. One monoclonal antibody generated against BoLA-NC3*00101 was highly specific. Unfortunately, due to failure to clone the NC3*00101- hybridoma, we no longer have an infinite source of this monoclonal antibody for NC3*00101. We eluted peptides from NC3*00101-transfected MHC-null K562 cells and identified peptides using liquid chromatography-mass spectrum (LC-MS) analysis. Analysis of peptide binding data using the SAS Proc mixed statistical program, suggested that the peptide EVTNQLVVL is a potential peptide ligand, which can be used to make tetramers for enumeration of antigen-specific leukocytes.