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Genomic mammary amyloid a sequence

a gene and amyloid technology, applied in the field of new isoforms of serum amyloid a, can solve the problems of limited saa production outside the liver and have been found, and achieve the effect of stimulating saa production and stimulating mucin 3

Inactive Publication Date: 2006-02-02
TRITON ALGAL INNOVATIONS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0014] According to the invention the novel colostrum associated SAA particularly the highly conserved region, (TFLK) also has an ability to stimulate mucin 3 (MUC3) production. Thus colostrum associated SAA may be used to treat and prevent enteric infections or other mucin inhibited disease states, such as traveler's diarrhea, infant diarrhea, necrotizing enterocolitis, urinary infections, and provide veterinary medicine for prevention of diarrhea in herd animals. The invention thus includes pharmaceutical compositions comprising a pharmaceutically effective amount of colostrum associated SAA peptide and a carrier to treat these and other diseases with similar pathology. Finally, other epithelial cell linings of mucosal surfaces such as nasopharynx, bladder etc., which produce mucins may also be treated with the pharmaceutical compositions of the invention to stimulate mucin production to prevent or treat infections associated therewith.
[0015] In yet another embodiment, the specificity of the promoter can be used to stimulate SAA production to aid in treatment of diseases associated with the teats or other mammary tissue of animals. For example, the colostrum associated SAA promoter is induced by prolactin. Thus one could administer prolactin or other colostrum SAA inducing agent to stimulate its production and cause increased MAA at the mammary tissue of said animal. Other features and advantages of the present invention will be better understood by reference to the drawings, detailed descriptions and examples that follow.

Problems solved by technology

However, SAA production outside the liver has been found, on a limited basis.

Method used

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  • Genomic mammary amyloid a sequence
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Examples

Experimental program
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Effect test

example 1

Comparative Analysis of SAA in Serum, Colostrum and Whey

[0169] The purpose of this study was to determine if SAA levels in colostrum and whey corresponded to serum SAA levels in mastitis symptomless and symptomatic cows.

[0170] Colostrum, whey and serum samples were obtained from a challenge model study in which cattle were vaccinated against a gram positive organism. Two sets of samples were utilized: one set (4 cows) from vaccinated animals that displayed clinical symptoms of mastitis, and the other set (5 cows) from vaccinated animals showing no clinical symptoms. Sample designations are shown below:

Vaccinates - Non Clinical (NC)Vaccinates - Clinical (C)NC Cow AC Cow ANC Cow BC Cow BNC Cow CC Cow CNC Cow DC Cow DNC Cow E

Whey / colostrum samples were obtained from the quarter displaying the clinical symptoms.

[0171] ELISA assays were conducted according to standard protocols, e.g., as described by McDonald et al. (J. Immunol. Meth. 144: 149-155, 1991), using rat anti-SAA (human)...

example 2

Evaluation of SAA in Colostrum and Subsequent Serial Samplings of Milk

[0174] The purpose of this study was to evaluate colostrum and subsequent serial milk samplings to determine SAA content. Samples were obtained from Holstein dairy cows at the University of Nebraska—Lincoln Dairy Research Facility. Samples of colostrum were taken at calving, and subsequent milk samples were taken twice weekly for three weeks. Samples from all four udder quadrants were pooled. Results are shown in Table 2.

TABLE 2SAA Levels in Colostrum and Milk SamplesCow IDSample DaySAA ug / ml83colostrumCalving184.883milk+40.283milk+70.083milk+110.083milk+140.083milk+180.083milk+210.0908colostrumCalving135.6908milk+42.6908milk+79.1908milk+118.2908milk+142.0908milk+182.1908milk+213.6961colostrumCalving364.6961milk+40.3961milk+70.5961milk+110.0961milk+140.0961milk+180.0961milk+210.0

[0175] The results show that SAA is elevated in colostrum of normal animals but in very low levels or not detected in normal milk samp...

example 3

Purification of SAA from Colostrum

[0176] The procedure set forth below can be used for purification of SAA from serum, plasma, milk or colostrum from any animal species. The procedure comprises two basic steps: the SAA is purified to approximately 20% purity by hydrophobic chromatography, then further purified to approximately 95% purity by SDS-PAGE and electro-blotting.

[0177] Approximately 30 ml of octyl sepharose CL-4B (Pharmacia #17-0790-01) was prepared by washing it with approximately 10 volumes (300 ml) water to remove any traces of ethanol. This may be done by washing the gel in a sintered glass funnel (coarse, funnel volume 600 ml) or by adding the water to the gel in a beaker, and then allowing the gel to settle before pouring off the water and rewashing the gel.

[0178] The final washes (2×40 ml) of the gel were with a solution of 0.5M ammonium sulfate.

[0179] Prior to use, the colostrum was allowed to set at 4° C. to allow the lipid layer to separate from the aqueous lay...

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Abstract

A genomic nucleotide sequence encoding Serum Amyloid A (SAA), isolated and purified from mammalian colostrum, is disclosed. Methods of use for the same in transgenic protocols is also disclosed.

Description

FIELD OF THE INVENTION [0001] 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 [0002] 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. [0003] 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. [0004] The acute phase response involves a wide variety of mediators, including cytokines, interleukins and tumor nec...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N9/00C07H21/04C12P21/02C12N5/06A61K38/00G01N33/53A61K47/04A61K47/10A61K47/26A61P1/00A61P1/04A61P13/12C07K1/14C07K14/47C07K16/18C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12N15/12C12P21/08C12Q1/68G01N33/566G01N33/577
CPCC07K14/4711A61K38/00A61P1/00A61P1/04A61P1/12A61P13/02A61P13/12
Inventor MCDONALD, THOMASLARSON, MARILYNNWEBER, ANNIKA
Owner TRITON ALGAL INNOVATIONS
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