bluuebunny/crossref_metadata_2025 huggingface dataset:
ALLOGENEIC REACTIVITY IN NORMAL MOUSE SERUM
Authors: Lynes Michael A. , Flaherty Lorraine , Michaelson James , Collins Jeffrey J. , Rinchik Eugene M.
Published: April 2007

Normal C57BL/6 (B6) mouse serum was tested in the direct cytotoxicity assay for specific reactivity against lipopolysaccharide (LPS)‐stimulated mouse spleen cells. Selective reactivity was found in weanling and adult serum against lymphoblasts from mice that express an antigen encoded by the H‐2Kk region of the major histocompatibility complex (MHC). Other strains, congenic with B6 at the MHC, did not exhibit the same alloreactivity. Serum from mice of a congenic strain being derived in our laboratory, which differs from B6 at two unlinked loci, Tla and nu, exhibited similar reactivity against the B6‐H‐2k LPS‐stimulated lymphoblasts, implying that a competent T cell compartment is not necessary for generation of this reactivity. Such reactivities may result from environmental stimulation of the immune system, from internal immuno‐regulatory controls, or from some combination of these immune stimuli
MOLECULAR GENETIC ANALYSES OF THE MAJOR HISTOCOMPATIBILITY COMPLEX IN PIG FAMILIES AND RECOMBINANTS
Authors: Chardon P. , Renard Ch. , Kirszenbaum M. , Gefotin Cl , Cohen D. , Vaiman M.
Published: April 2007

Five HLA probes, one corresponding to class I genes, two corresponding to distinct class II light chain genes, DRβ and DQβ, and two to class II heavy chain genes, DRα and DQα, were used to analyse the genomic DNA of the pig. Three informative SLA typed families and fourSLArecombinants were studied by Southern blot analysis. About 16 restriction fragments, generated by EcoR 1 or Hind III endonucleases, were revealed for each individual, either with the class I probe or the DRβ probe. The number of restriction fragments which hybridized with the other probes was generally lower. Several restriction fragment‐length polymorphisms were found and these segregated withSLADRβ‐ and DQα‐like genes are probably tightly clustered within theSLA‐D‐MLRregion.
CHARACTERIZATION OF A MONOCLONAL ANTIBODY THAT RECOGNIZES A DETERMINANT UNIQUE TO HUMAN HAEMOGLOBIN β‐CHAIN
Authors: Krco C. J. , Gorzynski T. J. , Beito T.
Published: April 2007

Methodologies are described for the production and characterization of monoclonal antibodies to human haemoglobin. Three monoclonal antibodies are described, two of which recognize distinct determinants on the α‐chain subunit. A third monoclonal antibody, Hb‐2d, recognizes a determinant expressed on human β‐chain. The Hb‐2d determinant is shared by human and baboon haemoglobins, but is not expressed by haemoglobins from beef, goose, pig, rabbit, sheep, dog, rat or mouse. Monoclonal antibody Hb‐2d will bind to haemoglobin A2 but not to foetal haemoglobin suggesting that δ‐ but not γ‐chain also expresses the Hb‐2d determinant. The results of testing a limited panel of human haemoglobin variants is presented.
INTERPLAY OF IMMUNOGLOBULIN G HEAVY CHAIN MARKERS (Gm) AND HLA IN PREDISPOSING TO SYSTEMIC LUPUS NEPHRITIS
Authors: Stenszky V. , Kozma L. , Szegedi G. , Farid N. R.
Published: April 2007

We have studied the distribution of IgG heavy chain markers (Gm) among 90 Hungarian patients with systemic lupus erythematosus (SLE) (55 of whom were also typed for HLA). This study confirms previously described increases in HLA‐B8 and DR3 in this condition. No difference in the distribution of Gm phenotypes was found between patients and 168 controls from the same geographical area. HLA‐B8/Gm homozygous individuals were, however, at greater risk for SLE (relative risk = 5.13) compared to B8+Gm heterozygotes or B8− individuals, irrespective of Gm phenotype. When patients with renal manifestation (n= 40) were compared to those without, the Gm phenotype 3; 5, 13 was found to be significantly increased (x2= 10.36, P < 0.0001, relative risk (RR) = 4.69). HLA and Gm increased additively the risk for renal manifestations in that for those patients who were both Gm3;5,13+ and HLA‐B8+, PR was 110, while it was 21.2 for Gm3;5, 13−/B8+, 7.9 for Gm3;5, 13+/B8− and 1.0 for Gm3;5, 13−/B8− patients. The study suggests that combined HLA and Gm typing can be used to identify SLE patients at high risk for manifesting renal abnormalities.
MURINE EMBRYONAL CARCINOMA CELLS EXPRESS CLASS I MHC‐LIKE ANTIGENS
Authors: Stern P. L. , Bereword N. , Bell S. M. , Thompson S. , Jones K. , Mellor A.
Published: April 2007

Previous work has concluded that murine embryonal carcinoma (EC) cells, the oncogenic stem cells of teratocarcinomas, do not express class‐I, H‐2K or D/L gene encoded products. Experiments using cell‐mediated lysis, serological and molecular biological approaches show that EC cells do express some major histocompatibility complex (MHC) class I or class I‐like molecules.
HETEROGENEITY OF THE EXPRESSION OF CLASS I AND II HLA ANTIGENS IN HUMAN BREAST CARCINOMA
Authors: Perez M. , Cabrera T. , Lopez Nevot M. A. , Gomez M. , Peran F. , Ruiz‐Cabello F. , Garrido F.
Published: April 2007

HLA class I and II antigen expression was studied in 19 cases of primary infiltrating ductal carcinoma of the breast. An indirect immunofluorescence technique was used on cryostat sections with monoclonal antibodies directed against HLA class I and II monomorphic determinants. Of the 19 cases studied, 17 were positive for class I antigen expression and two were negative. Class I HLA antigen expression was found to be clearly heterogeneous: in ten of these tumours more than 75% of the cells were class I positive; in two the percentage was decreased to between 50% and 75%; in five tumours it was less than 50%. With respect to class II HLA antigen expression, eight breast tumours were totally negative while two were strongly positive. (50‐75%) and the nine remaining cases were less than 25% positive.

In addition, radioassay techniques were employed to determine the presence of oestrogen and progestagen receptors. The distribution of these receptors was not correlated with HLA class I or II antigen expression, nor could any relationship be demonstrated between the degree of histological differentiation of the tumours and their invasiveness.
EQUINE LEUCOCYTE ANTIGEN SYSTEM. IV. RECOMBINATION WITHIN THE MAJOR HISTOCOMPATIBILITY COMPLEX (MHC)
Authors: Lazary S. , Dubath M.‐L. , Luder Ch. , Gerber H.
Published: April 2007

A case of recombination between the putative class I ELA antigen series and the strudure(s) governing mixed lymphocyte reactivity in an informative horse family is described. The results of serological typing, ‘lysostripping’ and mixed lymphocyte culture tests strongly suggest that the recombination took place between two loci and is not intragenic. An alloantigenic membrane structure, provisionally called B1, which does not belong to the known ELA series, was also involved in the cross‐over. The B1 antigen resembles the class II gene products of other species in two respects: it is not present on platelets, and doantiserum with specificity for B1 inhibits the stimulatory effect of B1‐carrying cells in mixed lymphocyte cultures. The B1 antigen does not follow the classical distribution, however, being expressed on both B and T lymphocytes. The finding of separate loci for the first series of ELA antigens and the MLR governing structure(s) demonstrates the similarity of the genetic organization of the horse MHC to that in other species.
ABROGATION OF LETHAL GRAFT‐VERSUS‐HOST REACTION DIRECTED AGAINST NON‐H‐2 ANTIGENS: ROLE OF Mlsa AND K/I REGION ANTIGENS IN THE INDUCTION OF UNRESPONSIVENESS BY ALLOIMMUNIZATION
Authors: Halle‐Pannenko O. , Pritchard L. L. , Festenstein H. , Berumen L.
Published: April 2007

A graft‐versus‐host reaction (GVHR) directed against DBA/2 non‐H‐2 antigens alone can be induced by grafting B10.D2 bone marrow and spleen cells intravenously to heavily irradiated, H‐2d compatible (DBA/2 x B10.D2)F1 adult mice. Under the experimental conditions used, only 0‐l0% of recipients survive, but the survival is greatly increased by donor alloimmunization, a few days prior to grafting, against host‐specific (DBA/2) non‐H‐2 antigens and non‐specific (foreign) H‐2 antigens. The increased survival is mediated by alloimmunization‐activated suppressor cells which can decrease the intensity of the immune reaction developed by normal B10.D2 cells both in vivo (GVHR) and in vitro (proliferative response measured in mixed lymphocyte culture, MLC). The present experiments were designed to explore the antigenic requirements for inducing suppression. The results showed that in GVHR the protective effect induced by donor alloimmunization against the specific non‐H‐2 antigens, which leads to 70‐80% survival, is due primarily, if not entirely, to immunization against Mls2 antigens. Results of MLC experiments confirmed this conclusion, showing that immunization against Mlsa antigens is sufficient to account for the suppressive effect induced by the specific immunization. In addition, they indicated that the nonspecific protective effect induced by donor alloimmunization against foreign H‐2 antigens, which leads to 20‐30% survival, is due to immunization against antigens encoded by the K and/or I region(s) of the H‐2 complex; immunization against D region encoded antigens alone has no effect.
GENETIC INFLUENCE ON THE SERUM LEVELS OF NATURALLY OCCURRING HUMAN IGG ANTIBODIES TO DIETARY ANTIGENS: QUANTITATIVE ASSESSMENT FROM A TWIN STUDY
Authors: Husby S. , Schultz Larsen F. , Petersen P. Hyltoft
Published: April 2007

Serum IgG antibodies to ovalbumin (OA) and beta‐lactoglobulin (BLG) were quantified by ELISA techniques in 22 monozygotic (MZ) and 24 dizygotic (DZ) healthy twin pairs. Antibody levels were comparable in the MZ and DZ groups both for anti‐OA and anti‐BLG antibodies. The genetic variance (ĜWT) was 0.167 for log IgG anti‐OA antibodies, and 0·173 for log IgG anti‐BLG antibodies, with heritability estimates of 0·44 and 0·37, respectively. No indication was observed of genotype–environmental interaction or differential environmental covariance for the log antibody levels in the MZ and DZ twins. The anti‐OA and anti‐BLG antibody levels in the same individual correlated only to a low degree. The levels of naturally occurring serum IgG antibodies are significantly influenced by genetic factors.
MURINE CANDIDIASIS: III. HOST INFLAMMATORY RESPONSES ARE REGULATED IN PART BY CLASS I MHC GENES
Authors: Ashman R. B.
Published: April 2007

Host responses to experimental Candida albicans infection in mice have been shown previously to be regulated by genes in the major histocompatibility complex (MHC). Results reported here show that at least part of this control can be mapped to class I MHC genes.
A REAPPRAISAL OF Mls GENETICS
Authors: Abe R. , Hodes R. J.
Published: April 2007

Although the minor lymphocyte stimulatory locus (Mls) system was originally described as a single‐locus, multiple‐allelic system, allelism among the genes which encode Mlsa, Mlsc and Mlsd has not been demonstrated formally. In this report, genetic linkage between genes encoding Mlsa and Mlsc was studied by testing responses of unprimed T cells, as well as Mlsa‐ and Mlsc‐specific T cell clones, to the progeny of (AKR/J × C3H/HeJ) F1× B10.BR [(Mlsa×Mlsc)F1×Mlsb] crosses. In addition, the configuration of Mls genes expressed by CBA/J (Mlsd) was evaluated by examining the response pattern of a panel of Mlsd‐specific clones to (CBA/J × B10.BR)F1× B10.BR stimulators. Results of these segregation analyses indicated that the Mls system is composed of at least two independent and unlinked genes which encode, respectively, the determinants identified as Mlsa and Mlsc.
THE RELATIONSHIP OF Mlsx TO Mlsc
Authors: Abromson‐Leeman S. R. , Laning J. C. , Crowell J. , Dorf M. E.
Published: April 2007

Among T cell clones with specificity for cow insulin and autologous class II MHC products, a significant number displayed interesting patterns of alloreactivity to non‐MHC antigens. Four clones are described in this report. One is a typical Mlsa‐reactive clone, while the other three proliferate to a variety of allogeneic spleen cells with reportedly different Mls phenotypes. These include PL/J stimulator cells, designated Mlsx all strains reported to be Mlsc, and several strains previously typed as Mlsa. Little is known about Mlsx except that it does not appear to be cross‐reactive with Mlsa. In this report, therefore, we attempt to investigate the reasons why these clones seem to be stimulated by a variety of different Mls phenotypes. Our conclusions are, first, that some of the strains previously typed as Mlsa may actually express a second Mls product, either c or x, in a manner analogous to the CBA/J strain (which expressess both Mlsa and Mlsc), and second, that Mlsc and Mlsx are cross‐reactive. In preliminary experiments, we investigate the genetic relationship between Mlsc and Mlsx by analysis of backcrosses, and the extent of cross‐reactive recognition of Mlsc and Mlsx by raising T cell clones which recognize one but not the other. Our preliminary conclusion is that Mlsc and Mlsx are cross‐reactive, but represent distinct gene products.
Mls AND THE HELPER T‐CELL REPERTOIRE I. Mls as a regulator of T‐helper cell activation
Authors: Hämmerling U. , Toulon M. , Palfree R. G. E. , Hoffmann M. K.
Published: April 2007

We provide evidence that the Mls reaction involves a broad cross‐section of the helper cell population. In addition to those cells reacting overtly to Mls stimulatory spleen cells, there is a second large population of helper cells that are affected by an Mls difference. This latent Mls effect is manifested by either synergy or antagonism in mitogen‐mediated, Ia‐dependent T‐cell activation, depending on Mlsa or Mlsb phenotypes of stimulator cells, respectively. Our data can be explained by attributing to Mls alleles the function of differential regulation of autoreactivity of T cells. The results show that the concept of the Mls exerting a negative signal deserves serious consideration.
EXPRESSION OF HUMAN CD4 BY TWO HUMAN‐MOUSE INTERLINEAGE HYBRIDS
Authors: Taylor G. M. , Morten J. E. N. , Morten H. , Dodge A. B. , Rldway J. C. , Jones P. M. , Harris R.
Published: April 2007

Two hybrid cell lines expressing human CD4 were prepared by fusing human B‐lymphoid cells with the mouse T‐lymphoma BW5147. Hybrid TF42 was derived from a human B‐lymphoblastoid line and TF53.1 from a human B‐ALL. Variants of these hybrids expressing or lacking CD4 were isolated by sorting cells stained with the monoclonal antibody (mAb) OKT4 on a fluorescence‐activated cell sorter (FACS). Cytogenetic, isoenzyme and DNA analysis confirmed the presence of human chromosome 12 in the CD4+ hybrids, and revealed that CD4 expression by TF42 was associated with multiple copies of this chromosome. Of seventy mAb recognizing human T‐cell antigens screened on the CD4+ and CD4− variants of the two hybrids, only mAb recognizing CD4 and Leu 8 reacted with the CD4+ cells. These hybrids should be useful in the preparation, screening and analysis of anti‐CD4 monoclonal antibodies, and in studies of CD4 epitopes recognized by HIV.
CYTOTOXIC T LYMPHOCYTE RECOGNITION OF HYBRID MHC CLASS I MOLECULES
Authors: Kanda T. , Macchi M. J. , LaPan K. , Darsley M. , Appella E. , Frelinger J. A.
Published: April 2007

We have utilized a group of MHC class I genes produced by in vitro recombination between Dp and Dd to study recognition of MHC class I molecules by cytolytic T cells (CTLs). Both polyclonal allo‐specific and H‐2‐restricted CTLs require that α1 and a2 of the target class I molecule be derived from the same haplotype for efficient killing. By using T‐cell lines we showed that within the bulk population there must exist a fraction of T cells which can recognize epitopes in al or α2. Critical residues for T‐cell recognition have been identified using these chimeric genes.
DIRECT HLA TYPING BY ROSETTING WITH IMMUNOMAGNETIC BEADS COATED WITH SPECIFIC ANTIBODIES
Authors: Hansen T. , Hannestad K.
Published: April 2007

Paramagnetic monodisperse polymer beads were coated with purified human hybridoma antibodies specific for polymorphic HLA determinants. The mAb‐coated beads bound specifically to cells from individuals with the relevant HLA type. The rosettes formed by beads and cells were isolated with a magnet. Rosette formation was evaluated microscopically and used as criterion for positive typing. By this rosette assay, fast and reliable HLA typing of whole blood was possible.
ISOLATION OF EPIDERMAL LANGERHANS CELLS
Authors: Schmitt D. A. , Hanau D. , Cazenave J.‐P.
Published: April 2007

Langerhans cells (LC) play an important role in the skin immune system. They are bone marrow‐derived and function as the only accessory and antigen‐presenting cells in the skin. Several techniques for enriching these cells have been devised, and four, including density gradient centrifugation, use of cell sorter, panning and immunomagnetic separation, are discussed. It is concluded that the most satisfactory method for isolation of LC is based on density gradient centrifugation and the most satisfactory for depletion of epidermal cell preparations for LC is based on the immunomagnetic principle.
THREE ANTIBODIES TO HUMAN INTESTINAL BRUSH BORDER MEMBRANE ARE CHARACTERIZED AS ANTI‐Lea
Authors: Dickson L. , Richman P. I. , Shaw M.‐A. , Donald A. S. R. , Swallow D. M.
Published: April 2007

In the course of raising monoclonal antibodies to human colonic and small intestinal epithelium, three were isolated which recognized epitopes on many colonic glycoproteins and many jejunal glycoproteins taken from some individuals, but not on the same glycoproteins from others. The specificities of these antibodies were examined by carrying out inhibitions of the binding of the antibodies to the jejunal glycoproteins by blood group substances and purified oligosaccharides. The glycoproteins were separated by electrophoresis and transferred to nitrocellulose strips which were then used for the inhibition experiments. The results indicated that the antibodies bind to Lea‐active structures. The conditions were determined for achieving red cell agglutination by two of the antibodies.
GROSS GENETIC DIFFERENCES AMONG SUBSTRAINS OF NZB MICE
Authors: Weiss S. , Uematsu Y. , D'Hoostelaere L. , Alanen A.
Published: April 2007

Substrains of NZB mice have been compared by Southern blot analysis using several probes. The restriction fragment length polymorphism of probes derived from the Igh‐V, Igk‐V, Tcrα‐C loci and of the long terminal repeat of the mouse mammary tumour virus revealed that NZB/BlLwPtIbm were grossly different from NZB/BlNJ and NZB/BlOla. Comparison with mouse strains of the Igk‐V haplotypes a and d suggested that NZB/BiLwPtIbm contain genetic material of the C58 mouse strain.
CLONING AND CHARACTERIZATION OF A POLYMORPHIC CLASS I MHC GENE IN THE AKR LYMPHOMA K36.16
Authors: Sim B. C. , Hui K. M.
Published: April 2007

Cancers are the result of somatic heritable changes in certain genes. The AKR leukaemia K36.16 has been extensively studied in our laboratory. When compared to normal AKR thymocytes, the K36.16 tumour cells do not express the H‐2Kk antigens and have an unexpected antigenic determinant that could be detected by anti‐H‐2Dd monoclonal antibodies. To understand the molecular mechanisms that could be responsible for these changes, we have compared the genomic composition of the class I MHC genes in the K36.16 tumour cells to that of normal AKR lymphocytes. A unique polymorphic 2.6‐kb Hind III fragment was detected in DNA obtained from the K36.16 tumour cells after hybridization with a 3′‐gene‐coding H‐2 probe. This fragment is not present in DNA of normal AKR lymphocytes. In an effort to further understand the mechanism underlying the nature of this MHC gene polymorphism, we have cloned and sequenced this Hind III fragment. When compared with the reported sequences of a number of mouse class I MHC genes, the nucleotide sequence of this polymorphic Hind III fragment is similar to that of a reported Tla gene.
TRANSFECTION OF MAJOR HISTOCOMPATIBLITY COMPLEX CLASS I AND CLASS II GENES CAUSES TUMOUR REJECTION
Authors: Ostrand‐Rosenberg S. , Clements V. K. , Thakur A. , Cole G. A.
Published: April 2007

Many human and mouse tumours do not express MHC class II antigens and have reduced levels of class I antigens. Because of the requirement for class I and/or class II antigen for antigen presentation to Th and Tc cells, these phenotypes may enable tumour cells to ‘escape’ the host's immune response. Experiments presented here are designed to assess the role of MHC class I and class II antigens in tumour immunity, and to overcome the MHC class I‐ or class II‐negative phenotype. When transfected with the syngeneic H‐2Db gene, the MHC antigen‐negative 402AX teratocarcinoma expresses high levels of H‐2Db antigen. 402AX/Db cells are rejected by MHC allogeneic and some WHC syngeneic 402AX‐susceptible mice, however the fully syngeneic strain of origin (129) remains tumour‐susceptible. Induction of MHC class I gene products on class I antigen‐negative embryonal carcinoma cells therefore increases tumour immunogenicity in some hosts, but not in the fully syngeneic mouse. In an attempt to enhance antigen presentation of tumour‐associated antigens to Th cells, MHC class I antigen‐positive SaI (KkDd) sarcoma cells were tranfected with syngeneic Aαk and Aβk genes to generate Iak‐expressing tumour cells. SaI/Ak cells are efficiently rejected by syngeneic A/J (KkDd) mice, while untransfected SaI cells are lethal. Induction of MHC class II antigen expression on the class I antigen‐positive SaI sarcoma therefore completely abrogates malignancy.
EVIDENCE THAT DERIVATION OF THE ADENOCARCINOMA LT85 PREDATED ESTABLISHMENT OF THE H‐2km2 MUTATION IN A C3HF COLONY OF MICE
Authors: Vogel J. M. , McMillan M. , Martin W. J. , Goodenow R. S.
Published: April 2007

The adenocarcinoma LT85 was chemically induced in a mouse from a C3Hf colony shown subsequently to be inbred for a gene conversion‐like mutation at the H‐2K locus, characterized by a clustered four nucleotide substitution in exon 3. The H‐2K phenotype of LT85, however, more closely resembles that of C3H rather than the mutant strain now designated C3Hf H‐2km2. We cloned and sequenced the H‐2K gene from this tumor to determine whether (i) LT85 might carry a tumor‐associated somatic reversion of the H‐2Kkm2 germline mutation or (ii) the tumor was induced in a mouse that was genetically H‐2k rather than H‐2km2. Our analysis confirmed that the LT85 H‐2K allele is identical throughout the entire coding region to C3H H‐2Kk. To exclude the possibility of a somatic reversion by recombination, we used an oligonucleotide probe corresponding to the region altered in H‐2k to show that the C3Hf genome lacks the necessary coding information to reverse the H‐2Kkm2 mutation through a sequence exchange with another class I locus. Since it is unlikely that multiple independent point mutations would account for restoration of this stretch of H‐2Kk sequences, the tumor was probably established in a mouse carrying a normal H‐2Kk allele, possibly at a point prior to the establishment of the H‐2Kkm2 mutation as a homozygous trait within the C3Hf colony.
MODULATION OF MHC GENE EXPRESSION IN HUMAN BREAST CARCINOMA CELLS BY HORMONES
Authors: Teh M. , Hui K. M.
Published: April 2007

Products encoded by the class I Major Histocompatibility Complex (MHC) genes serve as restriction molecules which enable T cells to generate an immune response to specific antigens. Recently, many investigators have demonstrated the importance of class I antigens in enabling the host to regulate tumor growth in vivo. In this report, we have studied the regulation of HLA genes by hormones in human breast cancer cell lines. Eight lines were studied. Using HLA locus‐specific DNA probes, the level of HLA‐A and HLA‐B specific mRNAs were found to be under‐represented in six of these cell lines when compared to an epithelial cell line derived from a normal lactating breast. Moreover, the expression of class I MHC mRNA in these cells correlated well with the level of chloramphenicol acetyltransferase (CAT) activity detected after the introduction of exogenous HLA‐CAT DNA‐constructs. It was also found that HLA expression in some of the breast carcinoma cell lines could be modulated by the addition of hormones. Hence, HLA mRNA expression in the cell line MCF‐7 was enhanced by the addition of estrogen; but was down‐regulated in the presence of dexamethasone. Conversely, for T‐47D cells, HLA expression was suppressed by progesterone. These results indicate that hormones could have an influence on the expression of HLA genes and may therefore indirectly be involved in the regulation of tumor growth by the host's immune system.
IL‐6 GENE POLYMORPHISM IN RHEUMATOID ARTHRITIS, PAUCIARTICULAR JUVENILE RHEUMATOID ARTHRITIS, SYSTEMIC LUPUS ERYTHEMATOSUS, AND IN HEALTHY DANES
Authors: Fugger L. , Morling N. , Bendtzen K. , Ryder L. , Andersen V. , Heilman C. , Pedersen F. Karup , Friis J. , Halbert P. , Svejgaard A.
Published: April 2007

The restriction enzymes MspI and Bg/II identify two different two‐allele restriction fragment length polymorphisms (RFLP) in the human IL‐6 genes of healthy Danes. Co‐dominant segregation was demonstrated for both marker‐systems and the test for Hardy‐Weinberg equilibrium showed no significant deviation from expectations. There is a strong correlation between the two marker systems. The two IL‐6 RFLP's were studied in Danish patients with rheumatoid arthritis, pauciarticular juvenile rheumatoid arthritis, and systemic lupus erythematosus. The frequencies of the MspI and Bg/II marker phenotypes did not differ between healthy controls and the three disease groups. No extra or missing DNA fragments were observed in the disease groups when compared with controls.
INFLUENCE OF RECOMBINANT INTERLEUKIN‐2 AND α‐ AND γ‐INTERFERONS ON THE INDUCTION OF NOVEL CLASS I ANTIGENS
Authors: Mitsuishi Y. , Fellous M. , Urlacher A. , Mayer S. , Tongio M. M.
Published: April 2007

Previous results obtained in our laboratory showed that novel class I antigens, closely related to HLA‐A (TM antigen related to HLA‐A9 and GO antigen related to HLA‐A24), were expressed on activated HLA‐A9 or HLA‐A24 peripheral blood lymphocytes (PBL), whatever activation factor was used [mitogenic stimulation (PHA, PWM, Con A), EBV transformation or alloactivation], but not on resting T and B lymphocytes. These antigens were also expressed on HLA‐A9 or HLA‐A24 common acute lymphoblastic leukaemias (CALL) and some ‘immature’ chronic lymphocytic leukaemias (B‐CLL), but not in hairy cell leukaemias (HCL), most of the B‐CLL, acute myeloblastic leukaemias (AML) and acute myelomonoblastic leukaemias (AMoL). In order to investigate the regulation mechanisms of these novel class I antigens, PBL and different leukaemic cell types were treated in vitro with recombinant IL‐2 (rIL‐2) and α and γ‐interferon (rIFN). Our results showed that without previous activation, but after culture with rIL‐2, TM and GO antigens could be induced in HLA‐A9 or HLA‐A24 PBL and enhanced in HLA‐A9 or HLA‐A24 B‐CLL cases, whereas no expression was found in HLA‐A9 or HLA‐A24 HCL, T‐ALL, AML or AMoL. After culture with rINFs, the expression of TM and GO antigens could be induced on HLA‐A9 PBL and all HLA‐A9 leukaemic cell varieties. Our results support the hypothesis that the expression of class I antigens is induced at an early stage of the cell cycle.
HLA‐B8,DR3 PHENOTYPE AND LYMPHOCYTE RESPONSES TO PHYTOHAEMAGGLUTININ
Authors: Modica M. A. , Cammarata G. , Caruso C.
Published: April 2007

Several reports have shown that HLA‐B8,DR3 positive subjects may display some changes in immune parameters when compared with HLA‐B8,DR3 negative ones and are prone to develop several immunological diseases. In the present study we have analysed the proliferative response to phytohaemagglutin (PHA) in HLA‐typed healthy subjects. A twin method was also employed to assess the role of genetic and environmental factors in the regulation of the response to the mitogen. It was not possible to demonstrate any difference in proliferative response to optimal doses of PHA between groups of subjects carrying or not carrying the HLA‐B8,DR3 phenotype. When suboptimal responses were studied, however, the results showed that ly‐mphocyte responses were significantly decreased in HLA‐B8,DR3 positive subjects compared with the negative ones. Moreover, the experiments performed with twins demonstrated that environmental factors were more important than genetic factors in the proliferative response to mitogen. The fact that the HLA‐B8,DR3 phenotype affects the suboptimal response to PHA although environmental factors are more important than genetic factors in the response to the mitogen seems of some interest. However, these results could be consistent with the high incidence of autoimmune disorders among HLA‐B8,DR3 positive individuals.
INSULIN‐DEPENDENT DIABETES MELLITUS SECONDARY TO CHRONIC PANCREATITIS IS NOT ASSOCIATED WITH HLA OR THE OCCURRENCE OF ISLET‐CELL ANTIBODIES
Authors: Larsen S. , Hilsted J. , Jakobsen B. K. , Svejgaard A. , Marner B. , Worning H.
Published: April 2007

We assessed HLA‐DR types and investigated serum samples for islet‐cell cytoplasmic antibodies (ICA) in 31 Danish patients with chronic pancreatitis. The antigen frequencies were compared with those in 1177 unrelated healthy Danish controls. Twenty patients had insulin‐dependent diabetes and 11 had normal intravenous glucose tolerance. No significant differences in the frequencies of DR3, DR4, or DR2 were found between patients with insulin‐dependent diabetes and patients with normal glucose tolerance or between any of these groups and controls. ICA were negative in all patients with chronic pancreatitis. It is concluded that the beta‐cell dysfunction in insulin‐dependent diabetes in chronic pancreatitis differs from that of classical insulin‐dependent diabetes.
THE NATURE AND ABUNDANCE OF HUMAN RED CELL SURFACE GLYCOPROTEINS
Authors: Anstee D. J.
Published: April 2007

Available evidence suggests that there are at least 30 different proteins at the red cell surface. These proteins vary in abundance from a few hundred copies/cell to more than a million copies/cell. Recently, a new Glycophorin gene (Glycophorin E) has been identified and sequenced. In this paper evidence is presented that a monoclonal anti‐M detects the product of the Glycophorin E gene in normal red cells.
SEROLOGICALLY DEFINED Rh DETERMINANTS
Authors: Tippett P.
Published: April 2007

Forty‐six numbered Rh antigens which are detected in serological tests are extant; antigens which were most recently numbered are of high or of low incidence. The identification of high incidence Rh antigens through studies involving cells with rare Rh phenotypes, including Rhnull cells, is described briefly. The serological investigations and family studies required to show that a low incidence antigen is an Rh antigen are illustrated by a description of the JAL (Rh48) antigen. Heterogeneity of polymorphic Rh antigens revealed by using monoclonal antibodies (MABS) is demonstrated by consideration of the Rh antigen D. A summary of results of serological tests shows that monoclonal anti‐D antibodies define eight D epitopes. Partial D antigens of the D categories are reassessed in terms of expression of D epitopes.
REPORT OF STUDIES ON MONOCLONAL ANTI‐IgG ANTIBODIES
Authors: Voak D. , Nilsson U.
Published: April 2007

Eleven monoclonal anti‐IgG antibodies were evaluated by ten laboratories for possible AHG reagent use by the ISBT/ICSH protocol. Three of the anti‐IgGs were selected by tests with weak Fya sensitized cells as this represents the best screening test. One of the IgM class anti‐IgGs (205) equalled ISBT/ICSH reference reagents (R3P and RIIIM) against weak anti‐D, ‐K and Fya sensitized cells and it reacted with all four subclasses of IgG. Thus, subject to satisfactory stability and extended trials with a wide range of weak antibodies, monoclonal anti‐IgGs for AHG reagent use are now a possible alternative to conventional rabbit anti‐IgG.
CHARACTERIZATION OF MHC ANCESTRAL HAPLOTYPES ASSOCIATED WITH INSULIN‐DEPENDENT DIABETES MELLITUS: EVIDENCE FOR INVOLVEMENT OF NON‐HLA GENES
Authors: Christiansen F. T. , Saueracker G. C. , Leaver A. L. , Tokunaga K. , Cameron P. U. , Dawkins R. L.
Published: April 2007

Insulin‐dependent diabetes mellitus (IDDM) is associated with several DR3‐ or DR4‐containing ancestral haplotypes (AHs). Using pulsed field gel electrophoresis (PFGE), long range maps of 35 haplotypes have been derived and classified. Two diabetogenic DR3‐containing AHs (8.1 and 18.2) possess deletions in the central non‐HLA region; these have not been found on non‐diabetogenic AHs tested to date. In addition, 8.1 and 18.2 also carry other deletions not found on other AHs. Three DR4 containing AH lack a Not 1 site, which may imply excision of an unidentified gene. These and other data suggest that deletions may be relevant to the pathogenesis of autoimmune disease, possibly through causing quantitative differences in autoimmune responses involved in IDDM. The MHC contains several regions of potential interest in relation to susceptibility to IDDM; these may explain the association with only certain DR3‐ and DR4‐carrying AH and DR3,4 heterozygosity in terms of cis and trans interactions. On the other hand, the class I1 region may be particularly important in protection.
EVIDENCE AGAINST EXTENDED DR3‐RELATED HAPLOTYPES IN GRAVES' DISEASE
Authors: Weetman A. P. , McCorkle R.
Published: April 2007

Three HLA‐DR3‐linked polymorphisms of the DPA, DPB and DRB genes, previously associated with myasthenia gravis or coeliac disease, have been examined in Caucasian patients with Graves' disease. The patients did not differ from healthy DR3 subjects, indicating an absence of any association of Graves' disease with specific DR3 subtypes.
HLA‐DR, DO AND DP TYPING USING PCR AMPLIFICATION AND IMMOBILIZED PROBES
Authors: Erlich H. , Bugawan T. , Begovich A. B. , Scharf S. , Griffith R. , Saiki R. , Higuchi R. , Walsh P.S.
Published: April 2007

A simple, rapid, and precise method of typing HLA class II polymorphism would be valuable in the areas of disease susceptibility, tissue transplantation, individual identification and anthropological genetics. Here we describe a method of analysing class II sequence polymorphism based on polymerase chain reaction (PCR) amplification and hybridization with oligonucleotide probes. One valuable property of sequence‐based HLA typing strategies, like oligonucleotide probe hybridization, is that they reveal how and where two alleles differ, not simply that they can be operationally distinguished. The nature and location of HLA polymorphisms appears to be critical in disease association studies and are likely to be important in tissue typing for transplantation. New alleles at the DRB1, DPB1 and DQB1 loci are likely to be identified as this technology is applied to more and more samples, particularly in non‐Caucasian ethnic groups. A new allele is uncovered as an unusual pattern of probe binding and then confirmed by sequencing. This pattern is observed because class II polymorphism is localized to specific regions and virtually all ‘new’ alleles have polymorphisms in the region of probe binding. Obviously, any new allele with a new polymorphic sequence in a region for which typing probes are not available would not be revealed by oligonucleotide typing.

With the PCR primers and probes described here, 7 DQA1 alleles,* 15 DQB1 alleles, 18 DPB1 alleles, and 32 DRB1 alleles are distinguished. Additional primers and/or probes can, of course, increase the allelic discrimination of oligonucleotide dot blot typing. These horseradish peroxidase (HRP)‐labelled oligonucleotide probes are stable (>2 years when stored at 4°C) and the typing system is simple and robust. Over 500 samples from the CEPH pedigrees (unpublished data; A. B. Begovich, et al., manuscript in preparation) and >1000 unrelated samples have been typed by this procedure. Although this dot blot/oligonucleotide hybridization procedure is a powerful and precise method of HLA class II typing, the complexity of the procedure increases as the number of probes required for analysis increases. The reverse dot blot method, based on an array of immobilized probes, allows the typing of individual samples in one single hybridization reaction. In this approach, a panel of unlabelled oligonucleotides are immobilized to a nylon membrane. The PCR product is labelled during the amplification reaction by using biotinylated primers and hybridized to the membrane. The presence of bound PCR product specifically hybridized to a given probe is detected using streptavidin‐HRP conjugates and either chromogenic or chemiluminescent substrates. This method offers the simplest and most rapid approach to the HLA typing of clinical samples.
HLA CLASS II NUCLEOTIDE SEQUENCES, 1991
Authors: Marsh S. G. E. , Bodmer J. G.
Published: April 2007

The HLA class II sequences included in this compilation are taken from publications listed in the accompanying paper, Nomenclature for factors of the HLA system, 1990 (Bodmer et al., 1991), and Nomenclature for factors of the HLA system, 1989 (Bodmer et al., 1990). Where discrepancies have arisen between reported sequences the original authors have been contacted where possible, and necessary amendments to published sequences have been incorporated into this alignment. Future sequencing may identify errors in this list and we would welcome any evidence that helps to maintain the accuracy of this compilation.

In the sequence alignments identity between residues is indicated by a hyphen (.). An unavailable sequence is indicated by an asterisk (*). Gaps in the sequence are inserted to maintain the alignment between different alleles showing variation in amino acid number.
ANCESTRAL HAPLOTYPES CARRY HAPLOTYPIC AND HAPLOSPECIFIC POLYMORPHISMS OF BAT1: POSSIBLE RELEVANCE TO AUTOIMMUNE DISEASE
Authors: Degli‐Esposti M. A. , Leelayuwat C. , Dawkins R. L.
Published: April 2007

The human BAT1 gene, located in the central MHC region (–170kb centrometric of HLA‐B), is polymorphic and the polymorphism correlates with MHC ancestral haplotypes. Allelic RFLP patterns have been assigned to several ancestral haplotypes and have been shown to be ‘haplotypic’ (i.e. found on all examples of the same ancestral haplotype) and in some cases ‘haplospecific’ (i.e. unique to one ancestral haplotype). The relevance of the BAT1 polymorphism to susceptibility to Myasthenia Gravis (MG) has been investigated. The frequency of the BAT1 B allelic pattern is increased in patients with MG (n= 16) compared to an equal number of control subjects. The increase is due to the association between MG and the 8.1 ancestral haplotype (HLA A1, Cw7, B8, BfS, C4AQ0, C4B1, DR3, DQw2).
DNA TYPING OF HLA‐DQ ALLELES BY GENE AMPLIFICATION OF DQA AND DQB VARIABLE EXONS: ANALYSIS OF DQA/DQB HAPLOTYPES
Authors: Pera C. , Delfino L. , Angelini G. , Longo A. , Ferrara G. B.
Published: April 2007

In the present report, we describe a DNA typing method that allows detection of all the polymorphic variants of DQA1 and DQB1 second exons. By the oligotyping procedure provided in this paper, we are able to identify 8 DQA1 and 13 DQB1 alleles and to type random individuals in any heterozygous combination. We provide the hybridization and washing temperatures for using either 32P labelled or non‐radioactive probes. The discrimination power of this procedure, compared to serological and cellular techniques, is remarkable. Therefore, this typing method finds perfect application in transplantation immunology and it will be very helpful to optimize the matching of unrelated donors before BMT. It is apparent from our results that despite the linkage disequilibrium present between DQ and DR loci, a DR specificity may frequently be associated to different DQ haplotypes. This is the case for DR4, DR7, DR8, DR9, and DR13 specificities.
ONE‐STEP MOLECULAR HLA‐DR PRESCREENING EMPLOYING A SET OF 14 SEQUENCE SPECIFIC OLIGONUCLEOTIDES IN A NON‐RADIOACTIVE TETRAMETHYLAMMONIUM CHLORIDE HYBRIDIZATION PROTOCOL
Authors: Sorg U. R. , Enczmann J. , Sorg R. V. , Wernet P.
Published: April 2007

A novel non‐radioactive protocol for molecular generic HLA‐DR typing is introduced, employing sequence specific oligonucleotides (SSOs) enzymatically 3′‐labelled with biotin‐14‐dATP via terminal deoxynucleotidyltransferase in a tetramethylammonium chloride hybridization procedure. The detection reaction is carried out, using streptavidine conjugated horseradish peroxidase which is bound to the SSOs, in combination with a light emitting detection system. Fourteen SSOs and one control SSO are employed for generic HLA‐DR typing in a one‐step protocol. In order to demonstrate the suitability of this procedure, 5 homozygous typing cell lines and samples of 11 pretyped individuals which include most serologically defined HLA‐DR specificities (DR1, 2, 3, 4, 11, 12, 6, 7, 8, 9,10, 52, and DR53) are analysed with the panel of 14 SSOs. The typing results show that this protocol, which avoids the use of radioisotopes, combines high specificity and easy handling. It also allows typing of poorly amplified samples because even after longer exposition times no false positive signals were observed and is particularly suitable for routine molecular HLA‐DR typing on the generic level. In addition it can easily be adapted to DP and DQ typing or DR subtyping.
DIALLELIC POLYMORPHISM MAY EXPLAIN VARIATIONS OF THE BLOOD CONCENTRATION OF MANNAN‐BINDING PROTEIN IN ESKIMOS, BUT NOT IN BLACK AFRICANS
Authors: Garred P. , Madsen H. O. , Kurtzhals J. A. L. , Lamm L. U. , Thiel S. , Hey A. S. , Svejgaard A.
Published: April 2007

Mannan‐binding protein (MBP) is a lectin which, upon binding to certain carbohydrates, activates the classical pathway of complement without the involvement of antibody or C1q. Deficiency of the MBP is associated with an opsonic defect and recurrent infections during early life. An amino acid substitution in the exon 1 at codon 54 in the MBP gene (GGC [glycine] to GAC [aspartic acid]) has been shown to be closely associated with low MBP concentration in Caucasoids. The gene frequency of the mutant allele in this population has been estimated at 0.13. In the study described here, we investigated the association between the mutant allele and MBP protein concentration in Eskimos from East‐Greenland and black Africans from the Baringo District in Kenya. The frequency of the GAC allele was identical in Eskimos and Caucasoids (0.13). No overlap with regard to MBP concentration between the genotypes was found in the Eskimos. In contrast, the Africans revealed a low frequency of the GAC allele (0.009). However, the median MBP protein concentration was approximately 5 times lower among the Africans than the Eskimos. In 12.6% of the Africans and in 2.5% of the Eskimos, MBP was undetectable. Thus, MBP deficiency is the most frequent immunodeficiency so far described. The high prevalence of MBP deficiency among healthy individuals indicates that MBP deficiency also confers some selective advantages. We advance the hypothesis that MBP deficiency is maintained in populations because MBP deficiency decreases the infectivity of some intracellular micro‐organisms which are dependent on opsonization.
COMPLEMENT COMPONENT C4 DEFICIENCIES AND GENE ALTERATIONS IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS
Authors: Fan Q. , Uring‐Lambert B. , Weill B. , Gautreau C. , Menkes C.J. , Delpech M.
Published: April 2007

Deficiency of complement component C4 is considered playing a role in the genetic predisposition for systemic lupus erythematosus (SLE). The purpose of this study was to characterize the genomic alterations of the C4 and CYP21 genes in 40 caucasoid patients with SLE by C4 allotyping and by RFLP analysis. Nineteen patients (47.5%) carried C4A null alleles and eight patients (20.0%) C4B null alleles. SLE patients had more frequent C4A null alleles (47.5%) than healthy individuals (20%) (X2= 10.75; P < 0.005). The commonest molecular alteration in the patients with C4A null alleles was a large gene deletion affecting both C4A and CYP21A genes. However, among the patients with C4A null alleles, 16.7% persons had no detectable C4A deletion. The non‐expression of C4A gene might be due to defects at various levels of gene expression (i.e. transcription and translation).

Among the patients with C4B null alleles, 62.5% persons had no detectable gene lesion, whereas 37.5% showed a C4B deletion including both C4B/CYP21A or C4B/CYP21B genes.

Duplication of the C4B gene was not rare in SLE patients, as we found 15.0% of the patients with a heterozygous C4B/CY21A gene duplication. The patients typed as having C4B gene homoduplication (B1,1) demonstrated two long C4B loci, whereas heteroduplication (B1,2) displayed two short loci, therefore the type of C4B gene duplication may be related to the gene length.

In conclusion, C4 deficiencies observed in 26 of the 40 SLE patients studied were very heterogeneous. In every case, the gene alteration affected both C4 and CYP21 genes.
AN HLA‐DR TYPING PROTOCOL USING GROUP‐SPECIFIC PCR‐AMPLIFICATION FOLLOWED BY RESTRICTION ENZYME DIGESTS
Authors: Westman P. , Kuismin T. , Partanen* J. , Koskimies S.
Published: April 2007

A simple PCR‐based protocol for HLA‐DR typing suitable for a routine practice is described. The method involves, first, a PCR amplification with seven different, group‐specific (DR1, DR2, DR4, DR7, DR9, DR10, and DR3+5+6+8) primer‐pairs, and second, typing of HLA‐DR allele more exactly in DR1, DR2, DR4, and DR3+5+6+8 groups by digestion of PCR products with restriction enzymes distinguishing different HLA‐DR types within each of the groups. Altogether 24 HLA‐DR alleles, or any combination of these, can be typed. The whole procedure, starting from a blood sample, can be carried out during a single working‐day. The method was tested by typing a set of homozygous cell lines, as well as a local panel previously typed by PCR/oligotyping. Also, 227 patients waiting for transplantation were typed to test the method in a routine setting. The results suggest that this kind of approach gives reliable HLA‐DR types and works well in the routine use.
QUANTITATIVE EXPRESSION OF HLA CLASS I MOLECULES IN ACUTE NON‐LYMPHOBLASTIC LEUKAEMIA CELLS
Authors: D'Alfonso S. , Savoia P. , Pitti G. , Peruccio D. , Pozzi C. , Crepaldi T. , Falda M. , Ficara F. , Resegotti L. , Richiardi P. Momigliano
Published: April 2007

The present study concerns a panel of 33 acute non lymphoblastic leukaemia (ANLL) patients, previously typed for HLA‐A,B serological specificities and including samples with a normal HLA‐A,B phenotype (3,4 detected specificities) as well as samples with missing and extra specificities. Samples were analysed at the protein and/or RNA level in order to verify whether the observed typing anomalies were due to a modified quantitative expression of class I molecules. The number of HLA‐A.B assigned specificities correlated significantly with the cell surface class I expression detected by indirect immunofluorescence using the monomorphic anti‐class I MoAb W6/32 (Spearman rank correlation test, P < 0.01) and with the amount of class I Heavy Chain (HC, P < 0.05) and beta‐2‐microglobulin (β2m, P < 0.05) evaluated by Western blot in whole cell extracts. The RNA analysis suggested a HC‐β2m coordinated down regulation at the mRNA level in a patient with no assigned HLA‐A,B specificities. Another patient with no detectable HLA‐A,B specificities showed a low expression selectively of the β2m protein. The results reported here demonstrate a heterogenous quantitative HLA class I expression in ANLL blasts, analogous to results reported for solid tumours.