|
Post by LymeEnigma on Apr 23, 2008 10:59:35 GMT -8
Variation in a Major Surface Protein of Lyme Disease Spirochetes ALAN G. BARBOUR,'* SANDRA L. TESSIER,' AND STANLEY F. HAYES2 Laboratory of Microbial Structure and Function,' and Electron Microscopy Section, Operations Branch,2 National Institute of Allergy and Infectious Diseases, Rocky Mountain Laboratories, Hamilton, Montana 59840 Received 21 February 1984/Accepted 27 March 1984 Two monoclonal antibodies (116831 and H5TS) differed in their indirect immunofluorescence reactivity when tested against 14 strains of Lyme disease spirochetes. Strains were bound by both antibodies, by H6831 or H5TS alone, or by neither. Western blot and immunoprecipitation studies revealed that the determinants of both antibodies were associated with abundant proteins with apparent subunit molecular weights of ca. 34,000 (34K-range proteins). The following results indicated that the 34K-range proteins were exposed on the surface of the spirochetes. (i) Antibody H6831 agglutinated the spirochetes; (ii) immune electron microscopy showed that the H6831 determinant was associated with the outer membrane; (iii) radiolabeled H6831 bound to live organisms; and (iv) proteases effectively removed the 34K-range proteins from intact cells. With their demonstrated variability and exposure on the surface, the 34K-range proteins may contribute to the serotype specificity of Lyme disease spirochetes. Full text: iai.asm.org/cgi/reprint/45/1/94
|
|
|
Post by LymeEnigma on Apr 23, 2008 11:01:05 GMT -8
Cross-reactive antigenic domains of the flagellin protein of Borrelia burgdorferi B. J. Lufta, Corresponding Author Contact Information, J. J. Dunnb, R. J. Dattwylera, G. Gorgonea, P. D. Gorevica and W. H. Schubacha a Department of Medicine, State University of New York, Stony Brook, NY 11794, USA b Department of Biology, Brookhaven National Laboratories, Upton, NY 11973, USA Received 28 September 1992; accepted 7 April 1993. ; Available online 10 December 2002. Abstract The p41 flagellin of Borrelia burgdorferi is the most common antigen recognized by serum of patients with Lyme borreliosis. This antigen shares amino acid homology, particularly in the amino and carboxy termini, with periflagellar antigens found in other microorganisms including Treponema pallidum. We cloned and expressed the p41 open reading frame in Escherichia coli and expressed it both as TrpE fusion and full-length unfused proteins. Also, we generated deletion constructs of various portions of the gene. Sera from patients with late Lyme borreliosis and secondary syphilis were used to identify the recombinant proteins by immunoblot analysis. Sera from 26 patients with Lyme borreliosis, 20 with secondary syphilis and 10 controls were used to identify cross-reactive domains of the B. burgdorferi flagellin. The variable region (amino acids 131–234) of the protein was recognized by 59% (15/26) of patients with late Lyme borreliosis compared to 30% (6/20) of patients with secondary syphilis and no (0/10) control patients. It appears that cross-reactive epitopes between B. burgodorferi and T. pallidum extend to the variable region of the flagellin. www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6VN3-47DKX5K-1C&_user=516213&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000023038&_version=1&_urlVersion=0&_userid=516213&md5=ac38551de85386f64410d247b23870ca
|
|
|
Post by LymeEnigma on Apr 23, 2008 11:02:16 GMT -8
Immunologic and structural characterization of the dominant 66- to 73-kDa antigens of Borrelia burgdorferi. Luft BJ, Gorevic PD, Jiang W, Munoz P, Dattwyler RJ. Department of Medicine, SUNY, Stony Brook 11794. The 66- to 73-kDa proteins of Borrelia burgdorferi are dominant immunogens and expressed in all strains of B. burgdorferi. The humoral response to these Ag occurs relatively early during the course of infection. Two-dimensional Western blot analysis of this group of Ag revealed them to consist of a tetrad of proteins with apparent molecular mass of 66, 68, 71, and 73 kDa. Furthermore, in this study we demonstrate the 66-kDa protein to be a potent inducer of lymphoproliferation in the patient immune to B. burgdorferi. Monospecific polyclonal antibodies and mAb demonstrate that each of these proteins was immunologically distinct. However, direct amino acid sequence of the 66- and 68-kDa Ag was almost identical and had a high level of sequence similarity to the GroEL heat-shock protein (Hsp60) of Escherichia coli and the 60-kDa immunodominant protein of Treponema pallidum. The amino terminal sequence of the 71- and 73-kDa proteins of B. burgdorferi was almost identical and these proteins had remarkable sequence similarity to the DnaK heat-shock protein of E. coli (Hsp70). It appears likely, therefore, that proteins related to the heat-shock family are potent immunogens of B. burgdorferi. PMID: 2016526 [PubMed - indexed for MEDLINE] www.ncbi.nlm.nih.gov/pubmed/2016526?ordinalpos=37&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum
|
|
|
Post by LymeEnigma on Apr 23, 2008 17:08:57 GMT -8
Relapsing FeverTick-Borne Relapsing Fever in the Northwestern United States and Southwestern Canada Mark S. Dworkin, Donald E. Anderson, Jr., Tom G. Schwan, Phyllis C. Shoemaker, Satyen N. Banerjee, Barry O. Kassen, and Willy BurgdorferAbstract: Records from 182 cases of tick-borne relapsing fever (TBRF) were reviewed. In confirmed cases, there was febrile illness, and spirochetes were identified on peripheral blood preparations. In probable cases, there were clinical features of TBRF and either the same exposure as a confirmed case or serological (indirect fluorescent antibody test and western blotting [WB]) evidence of infection with Borrelia hermsii. Sera also were tested for antibody to Borrelia burgdorferi. We identified 133 confirmed and 49 probable cases of TBRF. A Jarisch-Herxheimer reaction was reported in 33 (54.1%) of 61 cases for which this information was available. Most patients who had antibodies to B. hermsii were serologically positive for B. burgdorferi, and WB demonstrated false positivity of testing for B. burgdorferi. Thirty-five (21%) of 166 cases were unreported to public health authorities. In 52 cases, there were more than two relapses before the diagnosis. This study demonstrates that TBRF is underrecognized and underreported and may be falsely identified as Lyme disease.www.journals.uchicago.edu/doi/pdf/10.1086/516273More on relapsing fever Western blot: "Both C5-deficient and wild-type mice exhibited similar reactivity against Borrelial Ags with a 35-kDa protein as the major immunogen. Western blot for IgM reactivity in serum from postpeak C3-deficient mice (lane 4), C3 wild-type mice (lane 5), and against sham control fractions from uninfected mice (lane 6). Both C3-deficient and wild-type mice exhibited similar reactivity against Borrelial Ags with a 39-kDa protein as the major immunogen." www.jimmunol.org/cgi/content/full/167/6/3029View Western blot: www.jimmunol.org/cgi/content-nw/full/167/6/3029/F3
|
|
|
Post by LymeEnigma on Apr 23, 2008 17:14:35 GMT -8
SyphilisEvaluation of a Treponema pallidum western immunoblot assay as a confirmatory test for syphilis"The assay detected antibodies in syphilitic serum or plasma from dilutions of specimens incubated overnight with test strips. A test was considered positive when at least three of four major antigens having molecular masses of 15.5, 17, 44.5, and 47 kDa" Abstract: jcm.asm.org/cgi/content/abstract/30/1/115
|
|
|
Post by LymeEnigma on Apr 23, 2008 17:20:52 GMT -8
LeptospirosisImmunoreactive outer membrane proteins of Leptospira interrogans serovar Canicola strain Hond Utrecht IV R. Amutha*, P. Chaudhuri, Amar P. Garg*, P.S. Cheema & S.K. Srivastava "Results: The protein profile of SS fractions indicated the presence of three major bands of 41, 32 and 25 kDa and minor bands of 85 and 46 kDa. The SI fraction in serovar Canicola revealed the presence of 112, 93, 77, 43, 36, 29 and 22.5 kDa as major bands and minor bands of 102 and 53 kDa. In immunoblotting, the SS proteins of 41, 32 and 25 kDa and SI proteins of 112, 77, 36 and 22.5 kDa were detected to be major immunogenic proteins." medind.nic.in/iby/t06/i11/ibyt06i11p569.pdfAnother study: Human Leptospirosis Caused by a New, Antigenically Unique Leptospira Associated with a Rattus Species Reservoir in the Peruvian AmazonMichael A. Matthias1#, Jessica N. Ricaldi1,2#, Manuel Cespedes3, M. Monica Diaz4, Renee L. Galloway5, Mayuko Saito6, Arnold G. Steigerwalt5, Kailash P. Patra1, Carlos Vidal Ore7, Eduardo Gotuzzo2, Robert H. Gilman8, Paul N. Levett9*, Joseph M. Vinetz1* "...we determined whether a LipL32 cross-reactive protein might be present in “L. licerasiae” serovar Varillal strain VAR 010T by Western immunoblot using a rabbit anti-L. kirschneri serovar Grippotyphosa LipL32 polyclonal antiserum. As expected, a protein of ~32 kDa was seen in L. interrogans serovar Icterohaemorrhagiae strain HAI188 (Figure 4). Surprisingly, a single, well-defined protein of ~27 kDa, less than the expected molecular mass of this protein, was detected in “L. licerasiae” strain VAR 010T." www.plosntds.org/article/info:doi%2F10.1371%2Fjournal.pntd.0000213View comparative Western blots: www.plosntds.org/article/slideshow.action?uri=info:doi/10.1371/journal.pntd.0000213&imageURI=info:doi/10.1371/journal.pntd.0000213.g004
|
|
|
Post by LymeEnigma on Apr 23, 2008 17:56:13 GMT -8
How easy is it for a Western blot to indicate the wrong disease?
B. burgdorferi cross-reactions:
kDa 18: C. pneumonia 23-25: B. Hermsi, leptospirosis (band 25), Yersina, C. pneumonia (band 25) 35: Yersina, C. pneumonia 39: B. Hermsi 41: S. pallidum, L. interrogans, Yesirna, potentially all spirochetes 60: S. pallidum, E. coli, Bartonella, Staphylococcus, M tuberculosis, E. coli 70: HGE
Proteins that could be close enough possibly to get confused, if test or interpretation is sloppy:
kDa 18: S. pallidum (17 kDa) 30: C. pneumonia (29 kDa) 31: L. interrogans (32 kDa) 34: B. Hermsi (35 kDa) 41: C. pneumonia (40 kDa) 45: T. pallidum (47 kDa), 66: M. tuberculosis, E. coli (65 kDa)
I'm sure there are more....
|
|
|
Post by enochroot on Apr 23, 2008 22:15:57 GMT -8
Looked at my WBs but they don't have breakdowns on them I recall from back with Dr C, the only band his WB test lit up on, was 41
|
|
|
Post by LymeEnigma on Apr 24, 2008 8:09:59 GMT -8
Band 41 was the only one of mine to light up, when I had the Quest WB done; it was quite a while later that I had the Igenex test performed.
I'm just musing, here, but I'm wondering how many Igenex IND results could possibly be a lighting up of the next closest available band...? I wonder, with all testing facilities, how easy it is to read one of those test strips incorrectly ... and I wonder how many people have relapsing fever, leptospirosis, brucellosis, syphilis, or even a bad case of E. coli, and are being treated instead for Lyme....
|
|
|
Post by LymeEnigma on Apr 24, 2008 9:21:14 GMT -8
|
|
|
Post by LymeEnigma on May 16, 2008 9:20:10 GMT -8
Looks like some researchers have had the same idea.... Molecular Typing of Borrelia burgdorferi Sensu Lato: Taxonomic, Epidemiological, and Clinical Implications: cmr.asm.org/cgi/content/full/12/4/633
|
|
|
Post by enochroot on Jun 25, 2008 0:57:17 GMT -8
|
|
|
Post by LymeEnigma on Jun 25, 2008 9:55:26 GMT -8
Ugh ... the picture of those ticks at the top of that page is going to haunt me all morning! Yes, there does seem to be a huge amount of controversy surrounding that one little band. What it all boils down to is a positive response to 41 kDa indicates, most likely, that a spirochete is present in the body. Whether or not that spirochete is H. pylori, T. pallidum, B. burgdorferi, or some other illness-inducing pathogen requires further testing. There are also numerous non-disease-causing spirochetes, and any of those bearing sufficient flagillin can also cause a positive on band 41.
|
|
|
Post by enochroot on Jun 26, 2008 5:08:28 GMT -8
Gee that's a better answer than most Docs give yah The best one of the I.D.'s I saw said " band 41?...Lyme ?.... Harrumph! That can mean anything"
|
|
|
Post by LymeEnigma on Jun 26, 2008 7:57:32 GMT -8
That was pretty much the response I got from my ID doc: that "anything" could cause a positive 41 kDa. And the guy actually said it with conviction ... where being on high horses is concerned, in my humble opinion, Dr. Lipman took the cake.
Seemed to pull a record amount of information out of his arse, as well....
|
|