Identification of HLA-C Locus Products by One-Dimensional Isoelectric Focusing (1D-IEF)

Hajek-Rosenmayr, A; Doxiadis, Ilias

doi:10.1007/978-1-4612-3552-1_68

Cited by 5 publications

(7 citation statements)

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“…However, more detailed familiar analyses are still needed. The banding properties of these gene products differ from those of the HLA-A and HLA-B genes, in that more than one band per product is observed, as already shown by molecular cloning of the HLA-Cwl and Cw2 genes [19], and by ID-IEF [4,5]. On the other hand the definition of the bands corresponding to the HLA-C gene products is exclusively done by cor relation to serological results.…”

Section: Discussionmentioning

confidence: 99%

“…First attempts have been made for the characterization and definition of the HLA-C gene products by the meth odology presented here [4,5]. However, more detailed familiar analyses are still needed.…”

Section: Discussionmentioning

confidence: 99%

“…Biochemically, HLA-A3 shared by the parents was found as the basic subtype (A3.2). The maternal HLA-B39 antigen represents the acidic subtype B39.1, and HLA-A2 is the subtype A2.2 [3], In conjunction to data presented at the 10th Interna tional Histocompatibility Workshop on the banding properties of the HLA-C gene products [4,5], the respective protein bands are shown by a dotted line. However, the HLA-Cw6 banding pattern in not restricted to the members (M, C3, C4) with the haplotype d (A2,Bw57,Cw6) but is also observed in the family mem bers (F, C2, C5) with the haplotype b (A25,B37,C-), though these members were typed as HLA-C'blank\ In figure 2 the class I pattern of family DE shows the acidic HLA-A3 subtype (A3.1 ) on the maternal haplotype d (HLA-A3,B7,Cw7) which is not segregating.…”

Section: Preparation Of Samplesmentioning

confidence: 99%

“…Biochemical techniques for studying the HLA poly morphism have utilized primarily radiolabelled B-lymphoblastoid cell lines (B-LCL) [3][4][5]7] and/or mitogenstimulated peripheral blood lymphocytes (PBL) [7][8][9][10]. In addition, only very few family segregation data are available to date.…”

Section: Preparation Of Samplesmentioning

confidence: 99%

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HLA Class I Biochemistry: Definition and Frequency Determination of Subtypes by One-Dimensional Isoelectric Focusing and Immunoblotting

Frenz¹,

Doxiadis²,

Vögeler³

et al. 1989

Vox Sanguinis

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Previous biochemical studies of HLA class I gene products were based on the use of radiolabelled lymphoblastoid cell lines and/or mitogen-stimulated peripheral blood lymphocytes (PBL). We present here a nonradioactive methodology for defining HLA class I polymorphism on a small number of cells, such as 5x10^6 lymphocytes, the vitality of which is not a requirement in contrast to radiolabelling techniques, where1-2x10^6 PBL are needed. We analyzed 54 serologically typed families with a total of 261 individuals and 214 haplotypes by one-dimensional isoelectric focusing and subsequent immunoblotting, using a polyclonal anti-class I heavy chain serum. The banding patterns segregated within families and correlated with the serological results. A substantial number of HLA-A and HLA-B products appeared biochemically heterogeneous allowing the definition of subtypes and the calculation of their frequencies.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Preparation Of Samplesmentioning

confidence: 99%

Section: Preparation Of Samplesmentioning

confidence: 99%

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HLA Class I Biochemistry: Definition and Frequency Determination of Subtypes by One-Dimensional Isoelectric Focusing and Immunoblotting

Frenz¹,

Doxiadis²,

Vögeler³

et al. 1989

Vox Sanguinis

Self Cite

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“…The concentration of HLA-A1 in homozygous (F) or heterozygous (M, Cl-C4) configuration in this family is lower than the concen tration of the other HLA gene products, regardless of the other HLA class I types of the family members. Not only HLA-A and HLA-B gene products are found in the plasma of the individuals tested but also the recently decribed HLA-C gene products (HLA-Cw7 in figure 1 and HLACw3 and Cw7 in figure 2) [15,16].…”

Section: Treatment Of the Blotsmentioning

confidence: 99%

Typing for HLA Class I Gene Products Using Plasma as Source

Doxiadis¹,

Grosse‐Wilde²

1989

Vox Sanguinis

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Soluble HLA class I proteins have been found in serum or plasma of healthy and diseased individuals. Here we present evidence that these molecules can be readily used for determination of the HLA type by biochemical methods. Immunoprecipitation of the soluble class I gene products using monomorphic monoclonal antibodies coated to immunobeads and one-dimensional isoelectric focusing followed by immunoblotting represents a feasible and reproducible technique for typing. Analysis of these gene products in families (n = 12, with a total of 62 individuals) as well as in the population (n = 82) showed that all serologically defined antigens tested to date were present in plasma. A reference chart established primarily for the membrane-bound antigens could also be used for the soluble ones.

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HLA-C “blank” alleles express class I gene products. Biochemical analysis of four different HLA-C “blank” polypeptides

et al. 1989

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Monoclonal antibodies (mAbs) W6/32, HC10, and 4E were used to precipitate class I antigens from 21 selected individuals with at least one HLA-C "blank" allele. In 19 of these individuals, characteristic HLA-C banding patterns which could be precipitated by all three HLA class I mAbs were observed on one-dimensional isoelectric focusing gels--obviously the gene products of HLA-C "blank". At least four allelic HLA-C "blank" gene products with different isoelectric points could be discerned. All of them segregated with HLA-C "blank" haplotypes in informative families; two of them were associated with HLA-B51, one with HLA-B38, and one with HLA-B18. Reactivity of the HLA-C "blank" heavy chains with mAb W6/32 indicates that they are able to associate with beta-2 microglobulin, and hence are most probably expressed at the cell surface.

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Identification of HLA-C Locus Products by One-Dimensional Isoelectric Focusing (1D-IEF)

Cited by 5 publications

References 1 publication

HLA Class I Biochemistry: Definition and Frequency Determination of Subtypes by One-Dimensional Isoelectric Focusing and Immunoblotting

HLA Class I Biochemistry: Definition and Frequency Determination of Subtypes by One-Dimensional Isoelectric Focusing and Immunoblotting

Typing for HLA Class I Gene Products Using Plasma as Source

HLA-C “blank” alleles express class I gene products. Biochemical analysis of four different HLA-C “blank” polypeptides

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