Molecular cloning of the canine CD44 antigen cDNA

Milde, Kerstin F.; Alejandro, Rodolfo; Mintz, Daniel H.; Pastori, Ricardo L.

doi:10.1016/0167-4781(94)90111-2

Cited by 6 publications

(4 citation statements)

References 20 publications

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“…To determine the nucleotide sequence of the CD44 variant exons, polymerase chain reaction (PCR) was performed by employing first two primers, E3f and E16r, complementary to exon 3 and 16 in the dog CD44s. Primers correspond to the nucleotide positions 328-347 (E3f) and 748-767 (E16r) in the recently reported dog CD44s sequence (Milde et al 1994). All primers are listed in Table 1.…”

Section: Cloning Of Dog Cd44 Variant Exonsmentioning

confidence: 99%

“…However, the overall homology of the dog CD44 sequence is closer to human than to rodent. Unlike CD44s, which is 90% identical to human at the amino acid level (Milde et al 1994), the variant exons have a much lower identity (71%). The identity at the amino acid level of dog vs human CD44 variant exons are summarized in Table 2.…”

Section: Determination Of the Cd44 Variant Expression In Healthy Dog mentioning

confidence: 99%

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Expression of CD44 variant transcripts in dog lymphatic tissue

1994

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The gene of the CD44 cell surface glycoprotein consists of 20 exons. Ten exons known as variant exons are inserted by alternative RNA splicing between exon 5 and 16, thus generating diversity in the extracellular portion of the protein. We have cloned the dog cDNA homologue of the human variant exon region of CD44 and characterized its transcript expression in normal lymphatic tissues. Using PCR with primers complementary to regions contiguous to the insertion point and a variant exon, all ten variant exons of dog CD44 were detected in a high number of different isoforms. Transcripts containing the ten variable exons directly spliced to both sides of the insertion point were detected. We found an extensive usage of all variant exons from v1 to v10 in a high number of different combinations, some presenting a tissue-specific expression pattern. A similar complex profile of transcript expression was found in peripheral blood lymphocytes from rat and human.

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Section: Cloning Of Dog Cd44 Variant Exonsmentioning

confidence: 99%

Section: Determination Of the Cd44 Variant Expression In Healthy Dog mentioning

confidence: 99%

Expression of CD44 variant transcripts in dog lymphatic tissue

1994

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“…Canine CD44 is a 85-to 90-kDa protein expressed in lymph node tissue [15] and canine CD44-specific antibodies have been used in transplantation studies [21,22]. A monoclonal CD44 specific antibody directed against a CD44 mRNA-expressing [3] macrophage/monocyte cell line from a dog with malignant histiocytosis [27] bound to macrophages/monocytes, subsets of lymphocytes and epithelial cells.…”

Section: Introductionmentioning

confidence: 99%

Up-regulation of the hyaluronate receptor CD44 in canine distemper demyelinated plaques

Alldinger

Fonfara

Kremmer³

et al. 2000

Acta Neuropathol

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CD44 antigen (CD44), the principle cell surface receptor for hyaluronate, is up-regulated in the human demyelinating disease multiple sclerosis on fibrous astrocytes. As astrocytes are the main target cell of canine distemper virus (CDV), the consequences of a CDV infection on the CD44 expression and distribution in brains with spontaneous demyelinating canine distemper encephalitis (CDE) were of interest. Thirteen acute, 35 subacute, and 11 chronic plaques of nine dogs with immunohistologically confirmed CDE and brains of control dogs were included in the study. For light microscopy, 5-micron-thick serial sections were stained with H&E and incubated with monoclonal antibodies (mAbs) against CD44 and canine distemper virus nucleoprotein and polyclonal antibodies (pAbs) against glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP). For immunoelectron microscopy, 90-nm-thick sections were double stained with anti-GFAP and anti-CD44 mAbs to specify CD44-expressing structures. In controls, CD44 was diffusely distributed in the white matter and single meningeal cells exhibited a marginal expression of the antigen. In acute and more prominently in subacute demyelinating encephalitis, there was a plaque-associated up-regulation of CD44 which paralleled GFAP. In chronic demyelinating lesions, a reduction of CD44 associated with a loss of GFAP-positive astrocytes was noted. Additionally, in chronic plaques, CD44 was expressed on the cell membrane of perivascular mononuclear cells. Immunoelectron microscopically, in controls, CD44 was rarely demonstrated on astrocytic cell processes. In contrast, in brains with CDE CD44 was found on the cell membrane of broadened astrocytic cell processes. In summary, CD44 is up-regulated on astrocytes in the early phase of CDE and seems to represent a marker for the activation of immune cells in the late phase of the infection.

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“…Canine CD44 antigen cDNA has been cloned and Western blot analysis revealed a 85-90 kDa protein expressed in lymph node tissue (MILDE et al, 1994). So far, anti canine CD44 specific antibodies have been used in transplantation studies (SANDMAIER et al, 1990;SCHUENING et al, 1987) and it is known, that CD44 is expressed on a variety of immune cells (COBBOLD and METCALFE, 1994).…”

Section: Introductionmentioning

confidence: 99%

Characterization of a Canine CD44 Specific Monoclonal Antibody

Alldinger

Baumgärtner

Kremmer

et al. 1999

Journal of Veterinary Medicine Series A

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Monoclonal antibodies (mAbs) were generated against a CD44 mRNA expressing (RT‐PCR) macrophage/monocyte cell line (DH82) from a dog with malignant histiocytosis. The mAbs, that reacted with DH82 cells by FACS analysis were tested on formalin‐fixed, paraffin‐embedded tissues. Exclusively the incubation of DH82 cell pellets with mAbs from clone 2D10 resulted in a cell membrane associated immunoreaction. Immunoelectron microscopy specified, that the antibody bound exclusively to the cell membrane and processes of DH82 cells. The mAb was tested on a variety of normal canine tissues, including lymphoid, urinary, alimentary, respiratory, and endocrine organs, nervous tissues, liver, pancreas, skin, and muscles. Furthermore, tumour and inflamed tissues were tested for immunoreaction with the mAb. Immunohistologically, the 2D10 mAb reacted with macrophages/monocytes, subsets of lymphocytes, epithelial cells, and central nervous system white matter. FACS analysis of canine peripheral blood leukocytes showed, that a high proportion of lymphocytes and granulocytes were positive with this mAb. Western blot analysis revealed, that the 2D10 mAb bound to a protein with a molecular weight of about 85 kDa. The results of FACS and Western blot analyses, RT‐PCR, immunohistology and immunoelectron microscopy strongly suggest that the antigen detected by the 2D10 mAb is most likely the canine equivalent of human CD44, a cell bound hyaluronan binding proteoglycan.

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Molecular cloning of the canine CD44 antigen cDNA

Cited by 6 publications

References 20 publications

Expression of CD44 variant transcripts in dog lymphatic tissue

Expression of CD44 variant transcripts in dog lymphatic tissue

Up-regulation of the hyaluronate receptor CD44 in canine distemper demyelinated plaques

Characterization of a Canine CD44 Specific Monoclonal Antibody

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