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Feline leukemia virus (p27)

ANTISERUM: Custom Monoclonals International. Chris Grant ( ckgrantcmi@rcip.com). Mouse monoclonal antibody. Clone: PF12J-10A. isotype: IgG1k

SPECIFICITY ACCORDING TO MANUFACTURER: With Western blot this protein gives strong bands at about gpr80kD (glycosylated FeLV gag gene product, 10+p12+p27), then several bands representing assorted partial gag gene products, then a strong band at p28.5 (p27), and perhaps one or two weak bands >27kD (p27 degradation products).

REACTIVITY ACCORDING TO MANUFACTURER: Glycoprotein 27 (core protein) of feline leukemia virus. It has been reported to work on formalin-fixed and paraffin-embedded tissues.

WORKING DILUTION: 1/500. Pretreatment with steamer (citrate buffer, pH 6.0)

METHOD: LSAB 2-PO. 30 min. RT.

CELLS/TISSUES STAINED (feline tissues): Bone marrow: mainly megakaryocytes with diffuse cytoplasmic staining. Probably also erythroid and myeloid cells. Spleen: Mononuclear cells in red pulp. Dendritic-like cells pattern in germinal centers of lymphoid follicles. Kidney: No reaction. Small intestine: Strong reaction in crypt epithelium (diffuse) and some mononuclear cells deep in bottom of the lamina propria..

REFERENCES:

Kipar A, Kremendahl J, Addie DD et al. Fatal enteritis associated with coronavirus infection in cats. J Comp Path 119:1-14.

Kovacevic S, Kipar A, Kremendahl J et al. (1997) Immunohistochemical diagnosis of feline leukemia virus infection in formalin-fixed tissue. Eur J Vet Path 3:13-19.


Feline leukemia virus (p15e)

ANTISERUM: Custom Monoclonals International. Chris Grant (ckgrantcmi@rcip.com). Mouse monoclonal antibody. Clone: PF6J-2A1. Isotype: IgG2ak

SPECIFICITY ACCORDING TO MANUFACTURER: FeLV transmembrane, mapping towards the amino-terminal domain. This antibody gives two bands on Western blot at 20 and 15 kD. The former is the entire FeLV p15e, including the intracellular sequence, and the latter is the truncated p15e sheared from the cell membrane surface.

REACTIVITY ACCORDING TO MANUFACTURER: Feline leukemia virus transmembrane (envelope) protein p15e. It has been reported to work on formalin-fixed and paraffin-embedded tissues.

WORKING DILUTION: 1/200. Pretreatment with proteinase K.

METHOD: LSAB 2-PO. 30 min. RT.

CELLS/TISSUES STAINED (feline tissues): Bone marrow: No reaction. Spleen: Dendritic-like cells pattern in germinal centers of lymphoid follicles. Kidney: No reaction. Small intestine: No reaction.

REFERENCES:

Kovacevic S, Kipar A, Kremendahl J et al. (1997) Immunohistochemical diagnosis of feline leukemia virus infection in formalin-fixed tissue. Eur J Vet Pathol 3:13-19.

Kipar A, Kremendahl J, Addie DD et al. Fatal enteritis associated with coronavirus infection in cats. J Comp Pathol 119:1-14.


Feline leukemia virus (gp70)

ANTISERUM: Custom Monoclonals International. Chris Grant ( ckgrantcmi@rcip.com). Mouse monoclonal antibody. Clone: C11D8-C21. Isotype: IgG2Bk

SPECIFICITY ACCORDING TO MANUFACTURER: It recognizes amino-acids 214 to 218 in gp70 sequence. This antibody may weakly crossreact with amino-acid sequence located in intracellular portion of FeLV p15e transmembrane (envelope) protein to give the 20 kD band in Western blots. With this technique this antibody gives bands at 85, 70, 45 (weak) and 20 kD.

REACTIVITY ACCORDING TO MANUFACTURER: Feline leukemia virus glycoprotein 70. It has been reported to work on formalin-fixed and paraffin-embedded tissues.

WORKING DILUTION: Steamer at 1/500.

METHOD: LSAB 2-PO. 30 min. RT.

CELLS/TISSUES STAINED (feline naturally infected with FeLV):

Bone marrow: mainly megakaryocytes with diffuse cytoplasmic staining. Probably also erythroid and myeloid cells.

Spleen: Mononuclear cells in red pulp. Dendritic-like cells pattern in lymphoid follicles.

Kidney: Interstitial cells deep in the cortex (macrophage-like).

Small intestine: Interstitial cells in lamina propria and rare cells in villar epithelium.

REFERENCES:

Abkowitz JL, Holley RD, Grant CK (1987) Retrovirus-induced feline pure red cell aplasia. Hematopoietic progenitors are infected with feline leukemia virus and erythroid burst forming cells are uniquely sensitive to heterologous complement. J Clin Invest 80:1056-1063.

Elder JH, McGee JS, Munson M et al. (1987) Localization of neutralizing regions of the envelope gene of feline leukemia virus by using anti-synthetic peptide antibodies. J Virol 61:8-15.

Grant CK, Emisse BJ, Jarrett O et al. (1983) Feline leukemia virus envelope gp70 of subgroups B and C defined by monoclonal antibodies with cytotoxic and neutralizing functions. J Immunol 131:3042-3048.

Kovacevic S, Kipar A, Kremendahl J et al. (1997) Immunohistochemical diagnosis of feline leukemia virus infection in formalin-fixed tissue. Eur J Vet Pathol 3:13-19.

Nicolaisen-Strouss K, Kumar HPM, Fitting T et al. (1987) Natural feline leukemia virus variant escapes neutralization by a monoclonal antibody via amino acid change outside the antibody-binding epitope. J Virol 61:3410-3415.

Sheets RL, Pandey R, Klement V et al. (1992) Biologically selected recombinants between feline leukemia virus (FeLV) subgroup A and an endogenous FeLV element. Virology 190:849-855.

PDF File

IMAGES

Spleen, cat. Dendritic cells in lymphoid follicle have FeLV antigen. Immunoperoxidase-DAB for FeLV.


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