Serum amyloid a isoform from colostrum

Details Serum amyloid a isoform from colostrum Serum amyloid a isoform from colostrum

1999-10-22

2003-01-21

Carlson, Karen Cochrane (Department: 1653)

Chemistry: natural resins or derivatives; peptides or proteins;

Peptides of 3 to 100 amino acid residues

Separation or purification

C530S300000, C530S328000, C530S330000

Reexamination Certificate

active

06509444

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to the field of immunology and mammalian immune systems. In particular, the invention provides novel isoforms of serum amyloid A, which are found in colostrum.
BACKGROUND OF THE INVENTION
Several scientific or patent publications are referenced in this patent application to describe the state of the art to which the invention pertains. Each of these publications is incorporated by reference herein, in its entirety.
Mammals respond to tissue injury, trauma or infection by executing a complex series of biological reactions in an effort to prevent further tissue damage, to initiate repair of damaged tissue, and to isolate and destroy infective organisms. This process is referred to as the inflammatory response, the early and intermediate stages of which are referred to as the acute phase response.
The acute phase response involves a wide variety of mediators, including cytokines, interleukins and tumor necrosis factor. It also involves a radical alteration in the biosynthetic profile of the liver. Under normal circumstances, the liver synthesizes a range of plasma proteins at steady state concentrations. Some of these proteins, the “acute phase” proteins are induced in the inflammatory response to a level many times greater than levels found under normal conditions. Acute phase proteins are reviewed by Steel & Whitehead (Immunology Today 15: 81-87, 1994).
One of the massively induced acute phase proteins is Serum Amyloid A (SAA). SAA actually comprises a family of polymorphic proteins encoded by many genes in a number of mammalian species. SAAs are small apolipoproteins that accumulate and associate rapidly with high-density lipoprotein 3 (HDL3) during the acute phase of the inflammatory response. Most SAA isoforms are induced in response to inflammation; however, certain SAAs (e.g., human SAA4) appear to be constitutively expressed or minimally induced in the inflammatory response.
The liver has been considered the primary site of SAA production. However, SAA production outside the liver has been found, on a limited basis. For instance, expression of SAA mRNA has been reported in human atherosclerotic lesions and in human cultured smooth muscle cells and monocyte/macrophage cell lines (Meek et al., 1994; Urieli-Shoval et al., 1994; Yamada et al., 1996), and a unique isoform of SAA (SAA3) is produced by rabbit synovial fibroblasts (Mitchell et al., J. Clin. Invest. 87: 1177-1185, 1991). More recently, it was discovered that SAA mRNA is widely expressed in many histologically normal epithelial tissues, including tissues of stomach, intestine, tonsil, breast, prostate, thyroid, lung, pancreas, kidney, skin and brain neurons (Urieli-Shoval et al., J. Histochem. Cytochem. 46: 1377-1384, 1998). The role of SAA in such tissues has not been elucidated, nor has it been determined if the SAAs present in those tissues are the same isoforms as those found in serum, or if they represent additional isoforms of SAA.
SUMMARY OF THE INVENTION
According to one aspect of the invention, a Serum Amyloid A (SAA) protein is provided, which is isolated and purified from mammalian colostrum. In one embodiment, the SAA is isolated and purified from horse colostrum. Preferably, the horse colostrum SAA comprises SEQ ID NO:3 or SEQ ID NO:4 and one or more of a sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8.
In another embodiment, the SAA is isolated and purified from cow colostrum, and preferably comprises an N-terminal amino acid sequence which is SEQ ID NO:1. Alternatively, the SAA is isolated and purified from sheep colostrum and preferably comprises an N-terminal amino acid sequence which is SEQ ID NO:2.
According to another aspect of the invention, an isolated nucleic acid molecule that encodes a mammalian colostrum SAA is provided. The nucleic acid molecule may be a gene, cDNA or RNA and may be single-stranded or double stranded. In a preferred embodiment, the nucleic acid molecule comprises a sequence that encodes one or more of SEQ ID NO: 1-8.
According to another aspect of the invention, a population of synthetic oligonucleotides is provided, which includes sequences obtained by back-translating one or more of amino acid SEQ ID NOS: 1-8. One or more members of this population of oligonucleotides specifically hybridizes to a gene or cDNA that encodes a colostrum SAA.
According to another aspect of the invention, antibodies immunologically specific for one or more epitopes of colostrum SAA. Preferably, the antibodies are immunologically specific for at least one epitope of the colostrum SAA that distinguishes colostrum SAA from serum SAA.
According to another aspect of the invention, a process is provided for obtaining SAA from a mammal. The process comprises the steps of: (1) providing a sample of colostrum from the mammal, and (2) separating SAA contained in the sample from other materials contained in the sample, thereby obtaining the SAA from the mammal. SAA produced by this process is also provided.
Other features and advantages of the present invention will be better understood by reference to the drawings, detailed descriptions and examples that follow.


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