Distribution of the A Blood Group Activity on Lipid and Nonlipid Fractions of Pig Organs

Krötlinger, F.; Meyer, J.-N.; Thiele, O. W.

doi:10.1159/000207588

Cited by 3 publications

(6 citation statements)

References 6 publications

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“…Analysis of porcine lymphocyte gangliosides revealed two major compounds, NeuGc-GM 3 and NeuGc 2 -GD 3 . The knowledge on ganglioside pattern of porcine lymphocytes are today limited.…”

Section: Discussionmentioning

confidence: 99%

“…Charts B in Figs 6 and 7 shows the staining of the pig lymphocyte gangliosides. An intense staining of the NeuGc-GM 3 and NeuGc 2 -GD 3 gangliosides was observed in the post-perfusion serum from Immune electron microscopic studies of pig cells released from pig kidneys during perfusion…”

Section: Immunostaining By Sera From Patients Extracorporeally Xenopementioning

confidence: 96%

“…Figure 5A shows the negative ion nanoES-MS spectrum of the total acidic fraction. The peaks in the mass range m/z 1167.7 to 1279.9 were identified as NeuGc-GM 3 Figure 5B shows the CID spectrum of the molecular ion [M-H]of NeuGc-GM 3 with d18 : 1-n22 : 0 creamide. Y-fragmentation peaks (saccharide loss) are seen at m/z 944.6 (Y 2 , M minus NeuGc), m/z 782.5 (Y 1 , M minus NeuGc-Hex) and m/z 620.5 (Y 0 , M minus NeuGc-Hex -Hex).…”

Section: Characterization Of Pig Lymphocyte Gangliosidesmentioning

confidence: 99%

“…Based on the thin-layer mobility, degradation analysis and mass spectroscopy, it is concluded that the two major porcine lymphocyte gangliosides are NeuGc-GM 3 and NeuGc 2 -GD 3 , respectively. Quantitative evaluation ( Table 2) showed that the concentration of NeuGc-GM 3 and NeuGc 2 -GD 3 was 0.2 and 0.4 nmol ganglioside/ mg cell protein, respectively.…”

Section: Characterization Of Pig Lymphocyte Gangliosidesmentioning

confidence: 99%

“…In addition to this major xenoantigen barrier, there are several other antigen systems which have to be considered. Examples of such are the swine lymphocyte antigen (SLA) system [2], the pig blood group A/O system [3,4], and the Hanganutziu–Deicher (H–D) antigens [5]. The latter antigens are N ‐glycolyl‐neuraminic acid (NeuGc) terminated gangliosides that are widely distributed in mammalian species such as pig and monkeys but absent in humans and chickens [6].…”

Section: Introductionmentioning

confidence: 99%

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Release of pig leukocytes during pig kidney perfusion and characterization of pig lymphocyte carbohydrate xenoantigens

Magnusson

Månsson

Strokan³

et al. 2003

Xenotransplantation

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The Galalpha1-3Gal (alphaGal) antigen is considered the main xenoantigen in the pig to human species combination but other porcine antigens have to be considered such as the swine lymphocyte antigen (SLA), the blood group A/O and the Hanganutziu-Deicher (H-D) antigens. The H-D antigens are N-glycolyl-neuraminic acid (NeuGc) terminated gangliosides that are widely distributed in mammalian species but absent in humans. Upon exposure to a vascularized pig organ, the human recipient can be immunized by direct interaction with the pig tissue or/and by transfer of tissue/cells from the organ into the recipient. In the present work, we describe the release of cells from porcine kidneys upon perfusion and the expression of glycolipid based alphaGal, blood group A/O and H-D antigens in pig lymphocytes. Pig kidneys were flushed with 20 ml of NaCl or Lidocain containing 5000 U heparin, and thereafter perfused with 3000-ml perfusion solution and the cells released were counted and examined microscopically. Neutral glycolipid and ganglioside fractions were extracted from purified pig lymphocytes. The extracted components were characterized by thin layer chromatography, degradation and mass spectrometry. The expression of alphaGal and H-D epitopes on cells released from pig kidneys and purified pig lymphocytes were studied by immune electron microscopy. A total amount of about 300 x 106 leukocytes, mainly lymphocytes were released in the perfusate from the kidneys, of which about 100 x 106 cells were eluated in the 600 to 2400 ml perfusate fraction. Immunelectron microscopical analysis with Griffonia simplicifolia isolectin B4 showed staining of pig leukocytes and other cells, morphologically similar to endothelial cells, released in the perfusate. The purified porcine lymphocytes contained 930 microg neutral glycolipid (4.2 microg/mg cell protein) of which 95% was glycolipids with one to four sugar residues. Immunostaining of the neutral glycolipid fractions revealed alphaGal terminated compounds migrating in the five and 10 to 12 sugar regions and blood group A compounds in the six and eight sugar regions. Two major gangliosides NeuGc-GM3 and NeuGc-GD3 were found in the pig lymphocytes. In a patient extracorporeally xenoperfused with a pig kidney, an increased staining of both alphaGal terminated structures as well as the H-D reactive gangliosides were found in the post-perfusion serum samples. In summary, leukocytes, mainly lymphocytes are released from pig kidneys during perfusion which may contribute to immunization of human xenograft recipients.

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“…Analysis of porcine lymphocyte gangliosides revealed two major compounds, NeuGc-GM 3 and NeuGc 2 -GD 3 . The knowledge on ganglioside pattern of porcine lymphocytes are today limited.…”

Section: Discussionmentioning

confidence: 99%

Section: Immunostaining By Sera From Patients Extracorporeally Xenopementioning

confidence: 96%

Section: Characterization Of Pig Lymphocyte Gangliosidesmentioning

confidence: 99%

Section: Characterization Of Pig Lymphocyte Gangliosidesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Release of pig leukocytes during pig kidney perfusion and characterization of pig lymphocyte carbohydrate xenoantigens

Magnusson

Månsson

Strokan³

et al. 2003

Xenotransplantation

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Chemical characterization of porcine blood serum components with A and SLA activity

Wrenger

Thiele

Cwik

et al. 1982

Animal Blood Groups and Biochemical Genetics

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SummaryThe porcine A blood group substance is found in the serum as a Lipid and, additionally, in certain animals, as a glycoprotein. Swine lymphocyte antigens (SLA) occur in the serum only as glycoproteins. Heat treatment of the solid residue obtained by lipid extraction yielded a water‐soluble fraction with low protein content, high A activity, but no SLA activity. Poly(glycosy1)ceramides with SLA activity do not occur in the serum; poly(glycosy1)ceramides with A activity cannot be excluded. Desialylation of protein fractions has no effect on A and SLA activity. Both A and SLA activities of protein fractions are stable to mild alkaline hydrolysis thus indicating N‐glycosidic carbohydrate‐peptide linkages.

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Expression of carbohydrate xenoantigens on porcine peripheral nerve

Magnusson

Strokan²,

Svensson

et al. 2004

Xenotransplantation

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The total amount of acidic glycolipids (gangliosides and sulpholipids) was 9 mg/g tissue for both the blood group O and A nerves with sulphatides as the dominating components. Analyses of the glycolipid fractions showed strong expression of both the alphaGal and the HD antigens in nerves from both blood group A and O pigs. In addition, small amounts of blood group A antigens were expressed in nerves from blood group A pigs. Staining of neutral glycolipids from blood group A pigs using monoclonal antibodies reacting with A antigen having different core structures suggested that the A epitope expressed on pig ischiadicus nerves is based on the type 1 core chain structure. Light and electron microscopical studies on the alphaGal and HD-antigen distribution revealed that the neural cells were alphaGal antigen negative. Endothelial cells of blood vessels, and lymphatic and perineural cells expressed alphaGal antigen. Both endothelial cells and myelinized axons revealed positively labelled for the HD antigen.

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Distribution of the A Blood Group Activity on Lipid and Nonlipid Fractions of Pig Organs

Cited by 3 publications

References 6 publications

Release of pig leukocytes during pig kidney perfusion and characterization of pig lymphocyte carbohydrate xenoantigens

Release of pig leukocytes during pig kidney perfusion and characterization of pig lymphocyte carbohydrate xenoantigens

Chemical characterization of porcine blood serum components with A and SLA activity

Expression of carbohydrate xenoantigens on porcine peripheral nerve

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