Caprine blood groups. 2. The C,G, H,I, J,K, L,N, and Q systems

Nine blood group systems of goats were identified using 12 caprine reagents produced by absorption of alloimmune antisera. The caprine C blood group system, possibly homologous to the ovine C blood group system, was characterized by two reagents and shown to be controlled by three alleles,C ¹²,C ²⁵, andC ⁻. A more complex blood group system of goats, designated G, was identified using three reagents and shown to be controlled by six codominant alleles (G ^10.19.20,G ^10.19,G ^10.20,G ¹⁰,G ¹⁹,G ²⁰) and a recessive allele (G ⁻). A further seven one-factor two-allelic systems were identified by seven reagents. The nine genetic systems provided exclusion probabilities of 0.479, 0.492, 0.548, and 0.572 in Australian Angora, Dairy, Cashmere, and Texan Angora goat breeds, respectively. This work was supported by a grant from the Australian Stud Book, Alison Road, Randwick, New South Wales 2031, Australia.     

Role of the B-G-region antigen in the humoral immune response to the B-F-region antigen of chicken MHC

In the chicken MHC there exist two regions, designated F and G, which were separated by crossing-over. The F region contains genes controlling all functions characteristic of the MHC. So far only one gene has been assigned to the G region and it is responsible for the presence of an RBC antigen. When cross immunizing animals of the congenic lines CB and CC with erythrocytes, we have found that both F- and G-specific antibodies were produced. By using the recombinant haplotypes B ^R1 and B ^R2 we were able to dissociate the F from the G antigen and immunize with them separately. It was found that production of F antibodies required the copresence of the G antigen, whereas G antibodies were formed regardless of the presence or absence of the F-region antigen. It could be demonstrated that a prerequisite of the role of the G antigen with respect to the F antigen was the localization of both antigens on the same erythrocyte. Possible mechanisms underlying this phenomenon are discussed.Abbreviations used in this paper MHC major histocompatibility complex - RBC red blood cells - PBL peripheral blood lymphocytes - GVH graft-versus-host - MLR mixed lymphocyte reaction - i.v. intravenous - PBS phosphate-buffered saline - ME mercaptoethanol     

Catalase polymorphism in the red cells of horses*

Catalase zymograms performed on hemolysates of horse red cells revealed three phenotypes each composed of a single zone of activity designated F, M and S in decreasing order of migration towards the anode. It is proposed that these phenotypes are controlled by a pair of alleles, Cat^F and Cat^S, such that the genotypes Cat^FCat^F, Cat^FCat^S and Cat^SCat^S give rise respectively to phenotypes F, M and S. The effects of storage, heat, UV light and selected reagents on these phenotypes are discussed.     

The R-C-E-F blood group system of chimpanzees: Serology and genetics

Six chimpanzee alloimmune antibodies define 20 phenotypes of the R-C-E-F blood group system, the counterpart of the human Rh system. Of the several specificities of this system, the R^c constitutes the crucial link with human Rh since the reactions of some chimpanzee alloimmune anti-R^c sera with human red cells parallel those obtained with human anti-Rh_o reagents. Reciprocally, properly absorbed human anti-Rh_o sera detect R^c specificity on chimpanzee red cells. Tests with large panels of human monoclonal anti-D antibodies confirm the notion of shared epitopes between human alloantigen Rh_o(D) and chimpanzee alloantigen R^c.     

Human-type blood groups and Gm factors in gibbons (Hylobates)

Blood specimens from 69 gibbons (63Hylobates lar, 4Hylobates concolor, and 2Hylobates pileatus) were tested for human-type ABO, MN, and Rh blood groups. AmongH. lar, three phenotypes were noted in the ABO and MN blood groups respectively, but all fourH. concolor were grouped as AM. All group A gibbons were of subgroup A₁; subgroups A₂B and A₁₂B were observed at a low frequency in group AB gibbons. Le^b antigen was detected in about 30% of the red cell samples fromH. lar, but all the samples were negative for Le^a. All the gibbons tested had c(hr) antigen but no other Rh antigens (D, C, E, and e) in their red cells. Some selected blood samples fromH. lar were also tested for some other blood group antigens and for the Gm and Inv factors. The Jk^a antigen was detected in all the red cell samples tested, but the S, s, U, K, k, and Fy^a antigens were not. In the tests of plasma with anti-Gm (1),H. lar could be divided into two groups, i.e., Gm(1)^Gi and Gm(–1)^Gi; Gm(2), Gm(4), and Inv(1) were absent in the species.     

TheRT1.G locus in the rat encodes a Qa/TL-like antigen

A new antigenic system in the rat homologous to theQa/TL antigen system in the mouse has been characterized. It was detected by antibodies raised in donor-recipient combinations that were matched for theRT1. A, B, D, E loci in the major histocompatibility complex (MHC): (R11×BN)F₁ anti-BN.1L(LEW), (R18×BN)F₁ anti-BN.1L, and BN.1LV1(F344) anti-BN.1L. Absorption analyses using these antisera and a variety of inbred, congenic and recombinant strains identified three alleles,RT1.G ^a ,G ^b ,G ^c , of whichG ^c is a null allele. The strain distribution of these alleles was determined, using 37 strains of rats representative of all of the prototypic haplotypes and a number of congenic and recombinant strains. The use of the congenic and recombinant strains showed that theRT1.G locus was linked to the MHC and that the most probable gene order wasA-E-G. Testcross analysis showed that the map distance betweenA andG was 1.4 cM(4/285 recombinants). The RT1.G antigen has a heavy chain ofM _r 46 000 and is present on both T and B cells.     

Plasma protease inhibitor (PI) system in the laboratory opossum,Monodelphis domestica

Protease inhibitor (PI) polymorphism was observed in the laboratory opossum,Monodelphis domestica, by either one-dimensional acid polyacrylamide gel electrophoresis (PAGE; pH 4.6) or isoelectric focusing (pH 3.5-5.0) followed by immunoblotting with rabbit antiserum to human α₁-antitrypsin; but acid PAGE produced superior resolution of the PI proteins. Family studies demonstrated an inheritance of nine codominant autosomal alleles,PI ^D ,PI ^E ,PI ^F ,PI ^G ,PI ^H ,PI ^I ,PI ^J ,PI ^K , andPI ^M , and a population study revealed frequencies of 0.411, 0.010, 0.341, 0.034, 0.023, 0.071, 0.035, 0.020, and 0.055, respectively.     

A novel α-N-acetylgalactosaminidase family with an NAD+-dependent catalytic mechanism suitable for enzymatic removal of blood group A antigens

Abstract

Enzymatic removal of blood group A and B antigens from the surface of red blood cells to develop universal blood was a pioneering vision originally proposed more than 25 years ago. A great variety of enzymes, potentially suitable for enzymatic conversion of red blood cells, has been described since, but the process has not been economically viable because of the poor kinetic properties and low pH optimum of enzymes. Recently, the identification of two new families of bacterial glycosidases with enhanced kinetic properties for the removal of A and B antigens at neutral pH marked a milestone in the field of transfusion medicine (). Here we present a detailed structural analysis of Elizabethkingia meningosepticum a-N-acetylgalactosaminidase (NagA) shown to efficiently cleave the A antigen. NagA, a member of glycoside hydrolase (GH) family 109, employs an unusual catalytic mechanism involving NAD⁺. Comparison of the active-center structure with that of members of GH family 4 reveals a striking degree of structural similarity that allows the postulation of a common reaction mechanism and illustrates a beautiful example of convergent evolution.     

Carbohydrate moieties of rat MHC class I antigens

The rat major histocompatibility complex class I antigens RT1.A^u and RT1.E^u from the u haplotype and RT1.Aⁿ from the n haplotype were labeled with ¹⁴C-asparagine or with ³H-fucose, mannose, galactose, and N-acetylglucosamine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed complete removal of radioactivity from the sugar-labeled antigen heavy chains by digestion with glycopeptidase F, an enzyme that removes N-linked glycans completely. High performance liquid chromatography analysis of the tryptic digests of the mixed sugar-labeled and asparagine-labeled antigens demonstrated that all the sugar-labeled peptides were coincident with asparagine-labeled peptides. The Aⁿ antigen showed three glycopeptides, each of which had different amounts of sugar radioactivity. The antigens A^u and E^u showed two glycopeptides with different amounts of radioactivity but at identical positions in the two antigens. Antigen E^u had an additional glycopeptide with a lower amount of radioactivity. The positions of the glycopeptides from the A^u and E^u antigens were different from those of the Aⁿ antigen. The peptide profiles of the ¹⁴C-asparagine-labeled A^u and E^u antigens demonstrated distinct differences between the molecules. The results of this study show that: (a) all the glycans on rat class I antigens are N-linked, as they are on H-2 and HLA class I antigens; (b) there are compositional differences among the glycans in each of the three antigens; (c) the glycosylation pattern of the rat class I antigens is similar to that of the mouse class I antigens, which contain two or three glycans, in contrast to that of the human class I antigens, which contain only one glycan; and (d) the antigens A^u and E^u from the same haplotype are more closely related to each other than they are to the Aⁿ antigen.     

Red cell H-deficient,salivary ABH secretor phenotype of Reunion island. Genetic control of the expression of H antigen in the skin

Based on the genetic model proposing thatH andSe are two structural genes, we predicted that the red cell H-deficient, salivary ABH secretor phenotype should be found on Reunion island, where a large series of H-deficient non-secretor families have been previously described. Two such Reunion individuals are now reported. POU [A_h, Le(a–b+), secretor of A, H, Le^a and Le^b in saliva] and SOU [O_h, Le(a–b+), secretor of H, Le^a and Le^b in saliva]. Both are devoid of H -2-fucosyltransferase activity in serum. In addition, the preparation of total non-acid glycosphingolipids from plasma and red cells of POU revealed the type 1ALe^b heptaglycosylceramide and small amounts of the monofucosylated type 1 A hexaglycosylceramide. Both glycolipids possess an H structure probably synthesised by the product of theSe gene. No other blood group A glycolipids, with types 2, 3 or 4 chains, normally present in the presence of the product of theH gene, were found on red cells or plasma of POU.TheH,Se andLe genetic control of the expression of ABH and related antigens in different tissue structures of the skin is described in 54 H-normal individuals of known ABO, secretor and Lewis phenotypes; in one red cell H-deficient salivary secretor (SOU); and in one H-deficient non-secretor (FRA). Sweat glands express ABH under the control of theSe gene. Sweat ducts express ABH under the control of bothH andSe genes and Lewis antigens under the control ofLe and bothH andSe genes. Epidermis, vascular endothelium and red cells express ABH under the control of theH gene. The products ofH andSe genes are usually expressed in different cells. However, the results illustrate that in some structures, like the epithelial cells of sweat ducts, both the products ofH andSe genes can contribute to the synthesis of the same Le^b structure.     

Synthesis of prostaglandin E2 and prostaglandin F2α by toadfish red blood cells

Saline washed red blood cells of the toadfish convert [1-¹⁴C] arachidonic acid to products that cochromatograph with prostaglandin E₂ and prostaglandin F_2α. This synthesis is inhibited by indomethacin (10 μg/ml). Conversion of arachidonic acid to prostaglandin E₂ was confirmed by mass spectrometry. When saline washed toadfish red blood cells were incubated with a mixture of [1-¹⁴C]-arachidonic acid and [5,6,8,9,11,12,14,15,-³H]-arachidonic acid, comparison of the isotope ratios of the radioactive products indicated that prostaglandin F_2α was produced by reduction of prostaglandin E₂. The capacity of toadfish red blood cells to reduce prostaglandin E₂ to prostaglandin F_2α was confirmed by incubation of the cells with [1-¹⁴C] prostaglandin E₂.     

A model for genetic analysis of programmed gene expression as reflected in the development of membrane antigens.

The red blood cell antigen determined in mice by the H-2^b allele was detectable by hemagglutination at birth, H-2^b-early (bE), in strain $\text{C57BL}\text{10}$, whereas that determined by H-2^a not until 3 days later, H-2^a-late (aL), in strain A. Each F₁ individual was both aL and bE. Timing genes segregated in the F₂ and BC₁ generations to give H-2^a-early (aE) and H-2^b-late (bL) in combinations with aL and bE in apparent Mendelian ratios. In some strain combinations in which H-2 alleles and strain background had been interchanged, timing type was transferred along with H-2, whereas in others it was the background which seemed to be more important for the timing type. Moreover, no recombinant types were found in the F₂ and BC₁ generations of these lines. Lines originating from the (B10 × A)F₂ generation were selected for reversed parental-strain-type phenotypes under a full-sib mating regime. True breeding aE or bL lines were not realized until the ninth to eleventh inbreeding generation. We propose a remote-cis-effect model consisting of one temporal gene tightly linked with H-2 and at least two others with special cis and recognition properties linked with each other but not with H-2.     

Measuring individual inbreeding in the age of genomics: marker-based measures are better than pedigrees

Inbreeding (mating between relatives) can dramatically reduce the fitness of offspring by causing parts of the genome to be identical by descent. Thus, measuring individual inbreeding is crucial for ecology, evolution and conservation biology. We used computer simulations to test whether the realized proportion of the genome that is identical by descent (IBD_G) is predicted better by the pedigree inbreeding coefficient (F_P) or by genomic (marker-based) measures of inbreeding. Genomic estimators of IBD_G included the increase in individual homozygosity relative to mean Hardy–Weinberg expected homozygosity (F_H), and two measures (F_ROH and F_E) that use mapped genetic markers to estimate IBD_G. IBD_G was more strongly correlated with F_H, F_E and F_ROH than with F_P across a broad range of simulated scenarios when thousands of SNPs were used. For example, IBD_G was more strongly correlated with F_ROH, F_H and F_E (estimated with ⩾10 000 SNPs) than with F_P (estimated with 20 generations of complete pedigree) in populations with a recent reduction in the effective populations size (from N_e=500 to N_e=75). F_ROH, F_H and F_E generally explained >90% of the variance in IBD_G (among individuals) when 35 K or more SNPs were used. F_P explained <80% of the variation in IBD_G on average in all simulated scenarios, even when pedigrees included 20 generations. Our results demonstrate that IBD_G can be more precisely estimated with large numbers of genetic markers than with pedigrees. We encourage researchers to adopt genomic marker-based measures of IBD_G as thousands of loci can now be genotyped in any species.     

Allotypes of anti-alloantibodies

Rat antibodies directed at the two known rat light-chain allotypes, characterizing strains DA and Lewis, were trace-labeled with ¹²⁵I. With these reagents, the allotypes of alloantibodies DA anti-Lewis and Lewis anti-DA could be determined in a sandwich technique involving rat red cells coated with alloantibody and topped with radioactive anti-allotype. When rat red cells were doubly coated with matching alloantibody and the corresponding anti-alloantibody [produced in (Lewis × DA)F₁ hybrid rats], both radioactive anti-allotype reagents were fixed to a significant extent. This suggests that the anti-alloantibody is an immunoglobulin produced by the F₁ hybrid and not a complex between the originally injected alloantibody and antigen.     

Three new blood groups of rhesus monkeys

Three new blood group systems, called “T,” “U,” and “V,” have been identified in the rhesus monkey (Macaca mulatta). Each system consists of a single antigenic factor (blood group) detected by a monospecific alloimmune reagent that agglutinates erythrocytes. The antisera that detect these blood groups were obtained following a series of alloimmunizations and absorption fractionizations of the resulting antisera to produce operationally monospecific typing reagents. Analyses of family data indicated that each blood group was controlled by an autosomal dominant gene and that each system was independent of previously defined systems. With the addition of these new blood groups, we can identify 16 different blood group systems and well over one hundred million possible phenotypes in this species.     

The Cholesterol-Dependent Cytolysin Pneumolysin from Streptococcus pneumoniae Binds to Lipid Raft Microdomains in Human Corneal Epithelial Cells

Streptococcus pneumoniae (pneumococcus) is an opportunistic bacterial pathogen responsible for causing several human diseases including pneumonia, meningitis, and otitis media. Pneumococcus is also a major cause of human ocular infections and is commonly isolated in cases of bacterial keratitis, an infection of the cornea. The ocular pathology that occurs during pneumococcal keratitis is partly due to the actions of pneumolysin (Ply), a cholesterol-dependent cytolysin produced by pneumococcus. The lytic mechanism of Ply is a three step process beginning with surface binding to cholesterol. Multiple Ply monomers then oligomerize to form a prepore. The prepore then undergoes a conformational change that creates a large pore in the host cell membrane, resulting in cell lysis. We engineered a collection of single amino acid substitution mutants at residues (A370, A406, W433, and L460) that are crucial to the progression of the lytic mechanism and determined the effects that these mutations had on lytic function. Both Ply^WT and the mutant Ply molecules (Ply^A370G, Ply^A370E, Ply^A406G, Ply^A406E, Ply^W433G, Ply^W433E, Ply^W433F, Ply^L460G, and Ply^L460E) were able to bind to the surface of human corneal epithelial cells (HCECs) with similar efficiency. Additionally, Ply^WT localized to cholesterol-rich microdomains on the HCEC surface, however, only one mutant (Ply^A370G) was able to duplicate this behavior. Four of the 9 mutant Ply molecules (Ply^A370E, Ply^W433G, Ply^W433E, and Ply^L460E) were deficient in oligomer formation. Lastly, all of the mutant Ply molecules, except Ply^A370G, exhibited significantly impaired lytic activity on HCECs. The other 8 mutants all experienced a reduction in lytic activity, but 4 of the 8 retained the ability to oligomerize. A thorough understanding of the molecular interactions that occur between Ply and the target cell, could lead to targeted treatments aimed to reduce the pathology observed during pneumococcal keratitis.     

Immunogenetic polymorphism of lipoproteins in swine. 1. Four additional serum β-lipoprotein allotypes (Lpp2, Lpp4, Lpp5 and Lpp15) in the Lpp system1

Four additional swine serum lipoprotein allotypes are described. Specific anti-allotype reagents were obtained from alloimmune precipitating sera produced in lipoprotein-defined-type recipients immunized with normal sera and subsequently with lipoprotein fractions. Identification studies indicate that the four serologically defined low-density lipoprotein (LDL) variants, designated Lpp2, Lpp4, Lpp5 and Lpp15, are members of a previously described Lpp system. The individual specificities, Lpp2, Lpp4 and Lpp5, are determined by three co-dominant autosomal genes, Lpp², Lpp⁴ and Lpp⁵, respectively, whereas the common specificity, Lpp 15, is controlled by a complex of genetic information of the Lpp- and Lpp* genes, and by the two previously described alleles, Lpp¹ and Lpp³; Lpp15 occurs on the same molecule with respective individual specificity. The Lpp5 and Lpp15 antigens behave as a pair of alternative allotypic specificities. The double immunodiffusion test in agar was employed to demonstrate independent phenotypic expression of each allelic gene in the Lpp heterozygous animals, for the analysis of the immune sera, and for lipoprotein testing of 3305 sera. Marked differences in gene frequencies were found between the swine breeds tested. As a result of characteristic frequencies, only nine of 15 possible Lpp genotypes were found in the breeding herds tested; the remaining six genotypes were obtained from testcross matings.     

Caprine blood groups. 1. The B system

Twelve of 24 monospecific caprine reagents produced by absorption of alloimmune antisera identified a complex blood group system of goats which was designated B, based on the results of a small comparison test with ovine reagents. The frequencies of the 12 B factors differed significantly among the Australian Angora, Texan Angora, Cashmere, and Dairy goat breeds. Three of the antigens detected by the reagents were shown to be related as linear subtypes, designated Ba₁, Ba₂, and Ba₃, and inherited as alleles. The segregations of B factors in 80 sire groups involving 1086 offspring demonstrated that groups of B factors (phenogroups) segregated as products of allelic genes. This work was supported by a grant from the Australian Stud Book, Alison Road, Randwick, New South Wales 2031, Australia.     

Studies on blood groups in the japanese quail: The common antigens possessed by red blood cells and leukocytes,and their inheritance

Two alloantigens, Ly₁ and Ly₂, were detected with alloantisera made by immunization with leukocytes. These antigens were present on red blood cells, peripheral leukocytes and spleen cells and found to be controlled by the autosomal codominant alleles. The phenotypic frequencies of Ly₁ antigen in the three quail stocks, 1, 2, and 3, were 6.7, 0, and 100 percent, respectively, and those of Ly₂ antigen were 0 percent in stocks 1 and 2, and 7.1 percent in stock 3. It was suggested that Ly₁ and Ly₂ antigens might be associated with the system controlling A(QN₁) antigen which was originally detected by the natural alloantibody. However, it remains to be investigated whether these antigens are associated with the major histocompatibility complex (MHC) of the Japanese quail.