03253nas a2200421   4500000000100000008004100001260001300042653001200055653002600067653001800093653001100111653002300122653001800145653002100163653001000184653002400194653001100218653002800229653004800257653002100305653001200326653002700338653003400365653001000399100001900409700001400428700001600442700001400458700001500472700001600487700001500503245012100518856007100639300001100710490000600721520209000727022001402817       1995                            d  c1995 Nov10aAnimals10aAntibodies, Bacterial10aBase Sequence10aCattle10aCloning, Molecular10aCrohn Disease10aGenes, Bacterial10aGoats10aHeat-Shock Proteins10aHumans10aMolecular Sequence Data10aMycobacterium avium subsp. paratuberculosis10aParatuberculosis10aRabbits10aSequence Analysis, DNA10aSequence Homology, Amino Acid10aSheep1 aEl-Zaatari F A1 aNaser S A1 aEngstrand L1 aBurch P E1 aHachem C Y1 aWhipple D L1 aGraham D Y00aNucleotide sequence analysis and seroreactivities of the 65K heat shock protein from Mycobacterium paratuberculosis.  uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC170216/pdf/020657.pdf  a657-640 v23 a<p>Mycobacterium paratuberculosis is the causative agent of Johne's disease, a chronic enteritis in ruminants. It has also been implicated as a possible cause of Crohn's disease, an inflammatory bowel disease of unknown etiology. The mycobacterial 65K heat shock proteins (hsp-65K) are among the most extensively studied mycobacterial proteins, and their immunogenic characteristics have been suggested to be the basis for autoimmunization in chronic inflammatory diseases. In this context, we isolated and sequenced the hsp-65K-encoding gene from our M. paratuberculosis PTB65K genomic library. A high degree of identity was found between the open reading frame (ORF) of the PTB65K gene and those of Mycobacterium tuberculosis (89.6%), Mycobacterium leprae (86.6%), and Mycobacterium avium 18 (98.8%). The amino acid sequence alignment of the PTB65K protein with the hsp-65K homologs revealed that the M. tuberculosis and M. leprae proteins each differed by 36 amino acid residues and that the M. avium 18 protein differed by 8 residues. We also investigated the humoral immune responses of animals with Johne's disease and patients with Crohn's disease against the recombinant PTB65K antigen. Immunoblot analysis showed that sera from only 3 of 10 clinically ill and 5 of 25 subclinically ill cows reacted with PTB65K. In addition, sera from two of two sheep and one of two goats with clinical symptoms of Johne's disease also reacted with PTB65K; 0 samples from 10 normal cows reacted. In humans, sera from 7 of 13 patients with Crohn's disease, 3 of 4 with tuberculosis, 5 of 6 with leprosy, 5 of 12 with non-inflammatory bowel disease, and 0 of 4 with ulcerative colitis reacted with the recombinant PTB65K antigen. These results indicate that this PTB65K heat shock protein is uninformative when used for serodiagnosis of Johne's disease in animals. However, in humans, the high intensity of antibody reactions of some sera from Crohn's disease patients compared with that from noninflammatory bowel disease patients showed a positive correlation with mycobacterial diseases.</p>  a1071-412X