Veterinary Immunology and Immunopathology, 34 (1992) 235-243 Elsevier Science Publishers B.V., Amsterdam

235

Cross-reactivity of monoclonal antibodies to bovine immunoglobulins with immunoglobulins of other species D. Henning and K. Nielsen Agriculture Canada, Animal Diseases Research Institute, NEPEAN, P.O. Box 11300, Station H, Nepean, Ont., K2H 8P9, Canada (Accepted 13 February 1992 )

ABSTRACT Henning, D. and Nielsen, K., 1992. Cross-reactivity of monoclonal antibodies to bovine immunoglobulins with immunoglobulins of other species. Vet. Immunol. Immunopathol., 34: 235-243. Monoclonal antibodies (Mabs) to bovine immunoglobulin heavy chain of the four major isotypes 7~, 72, a, # and the light chains (combined x and 2 ) were produced and found to cross-react in enzymelinked immunoassay (ELISA) with immunoglobulins of some other animal species despite the discrete specificity associated with an antibody derived from a single clone. This cross-reactivity, particularly amongst ruminants, could be utilized in serological testing for the diagnosis of disease in these species. For example, Mabs produced against bovine immunoglobulin light chain cross-react with bison immunoglobulin light chain and were used successfully in serological testing as the secondary detection antibody in an indirect ELISA for the diagnosis of Brucella abortus in bison herds in northwestern Canada. ABBREVIATIONS ELISA, enzyme-linked immunoassay; HRPO, horseradish peroxidase; Mabs, monoclonal antibodies; RT, room temperature.

INTRODUCTION

Cross-reactivity of immunoglobulin class does occur between species related phylogenetically. Bovine immunoglobulins share structural (Curtain and Fudenberg, 1973), biochemical (Conde et al., 1975) as well as antigenic characteristics (Aalund, 1968; Feinstein and Hobart, 1 9 6 9 ) w i t h other ruminants. Most studies on ruminant species have been done on sheep, goats and Correspondence to: D. Henning, Agriculture Canada, Animal Diseases Research Institute, NEPEAN, P.O. Box 11300, Station H, Nepean, Ont., K2H 8P9, Canada.

© 1992 Elsevier Science Publishers B.V. All rights reserved 0165-2427/92/$05.00

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D. HENNING AND K. NIELSEN

cows; however, Butler ( 1983 ) also reported cross-reactivity with non-ruminant species using polyspecific antiserum in an immunodiffusion test. Monoclonal antibodies (Mabs) are being used extensively as diagnostic reagents in veterinary medicine because they are permanently available, reproducible, specific antibodies. They are often utilized in the enzyme-linked immunosorbent assay (ELISA), a simple and sensitive immunotest used for the diagnosis of infectious disease. The initial cost of producing a specific usable Mab is very high; however, once developed and in use, further costs are minimal. It would be cost prohibitive to produce Mabs to each immunoglobulin isotype of every species of animal. Mabs to bovine immunoglobulin isotypes and bovine light chain were produced in mice and conjugated with the enzyme horseradish peroxidase ( H R P O ) , for use in an indirect ELISA for the diagnosis of Brucella abortus infection in cows. The cross-reactivity of these M a b - H R P O conjugates with 24 different animal species was examined by direct enzyme immunoassay. Although testing sera from different species using anti-bovine M a b - H R P O conjugates as detection reagents has been done to a limited extent, some data are presented to demonstrate how Mabs produced against bovine species of immunoglobulin have been used in testing for infectious disease in other species using an indirect ELISA. MATERIALS AND METHODS

Serum Several serum samples were collected from 24 different animal species including cow, yak, bison, buffalo, sheep, goat, reindeer, elk, caribou, camel, pig, human, horse, zebra, kangaroo, wallaby, rabbit, guinea pig, rat, mouse, chicken, turkey, goose and duck. These species are classified in Table 1. The serum samples (at least five) were pooled and the albumin was removed from the serum samples by precipitation of the globular proteins with a m m o n i u m sulfate (45% saturation). The resultant preparations, containing the immunoglobulins, were dialyzed against 0.15 M NaC1 overnight at 4 ° C. The protein concentration was estimated on the dialyzate by ultraviolet spectrophotometry at 280 n m using z~ litTl%cm-- 13.7 and concentrations were adjusted to 1 mg ml - 1 Mabs to bovine immunoglobulins Mouse Mabs against bovine L-chain, IgGL, IgG2, IgM and IgA were prepared by a minor modification of the method of Kennet et al. (1978 ).

Galliformes Galliformes Anseriformes Anseriformes

-

Macropodidae Macropodidae Leporidae Caviidae Muridae Muridae

Marsupialia Marsupialia Lagomorphia Rodentia Rodentia Rodentia

Aves

Bovidae Bovidae Bovidae Bovidae Bovidae Bovidae Cervidae Cervidae Cervidae Camelidae Suidae Huminidae Equidae

Artiodactylia (sub-order Ruminants) Ruminants Ruminants Ruminants Ruminants Ruminants Ruminants Ruminants Artiodactylia Primates Perissodactyla

Mammalia

Family

Order

Class

Common name

Cow Yak Bison Buffalo Sheep Goat Reindeer Elk Caribou Camel Pig Human Horse Zebra Kangaroo Wallaby Rabbit Guinea pig Rat Mouse Chicken Turkey Goose Duck

Taxonomic name

Bos taurus Pocphagusgrunniens Bison bison Bubalus bubalus Ovis aries Capra hircus Rangifer tarandus Cervis canadensis Rangifer Camelidae camelus Sus scrofa Homo sapiens Equus caballus Equus zebra Macro pus Petro gale Oryctolagus cunicuh" Cavia porcellus Rattus rattus Mus musculus Gallus gallus Meleagris gallopavo Anser albifrons Anserplatyrhynochos

12 13 11 12

100 68 100 54 100 68 37 64 53 7 8 17 10 7 12 9 11 11 10 8

L-chain

40 33 44 38

100 100 58 42 100 44 44 45 73 42 36 51 34 35 38 42 38 35 31 35

IgM

Percent reactivity

5 3 3 5

100 5 13 5 60 41 3 I00 3 3 7 5 5 7 5 5 5 3 5 7

IgG~

8 8 10 10

100 7 12 8 43 7 8 8 8 7 10 10 7 22 8 8 8 8 8 7

IgG2

46 58 71 56

100 100 59 94 100 47 47 56 94 50 57 68 85 59 73 47 73 62 58 88

IgA

Percent cross-reactivity of Mabs to a bovine immunoglobulinlight chain and heavy chain of each of the four major isotypes of animal species tested

TABLE 1

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Cross-reactivity of monoclonal antibodies to bovine immunoglobulins with immunoglobulins of other species.

Monoclonal antibodies (Mabs) to bovine immunoglobulin heavy chain of the four major isotypes gamma 1, gamma 2, alpha, mu and the light chains (combine...
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