The Western Blot, Lyme, Relapsing Fever, etc.

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From IgeneX
www.igenex.com/lymeset2.htm:

The IgG Western Blot is a sandwich-type immunoassay performed in a manner that allows visualization of the patient's antibodies. It is a qualitative test and is generally more sensitive and specific than the ELISA. This test must be used if the Lyme IgG/IgM antibody serology is equivocal or positive. The somewhat-specific Lyme antibodies of importance are against the following molecular weights of the B. burgdorferi antigens: 23-25 kDa (Osp C); 31 kDa (Osp A); 34 kDa (Osp B); 39 kDa; 41 kDa; and 83-93 kDa7. "kDa" is the abbreviation for "kilodalton," which is used for molecular weight designations. "Osp" refers to outer surface protein of the bacteria.
....
A positive IgG result with clinical history may be indicative of Lyme disease. Patients with other spirochetal disease and/or who test positive for rheumatoid factor or Epstein Barr virus may have cross-reacting antibodies. A positive response in this, as in any antibody assay, indicates sensitization, not necessarily active disease. 12
....
The antibody specificities of importance for the IgM blot are similar to those for the IgG blot (with the exception of 83-93 kDa, which is still being investigated for significance). The CDC/ASTPHLD criteria for a positive result are two of the following three bands: 23-25 kDa (Osp C); 39 kDa; and/or 41 kDa.8,9 IGeneX adds the 31 kDa (Osp A), and/or the 34 kDa (Osp B) to the criteria,10,12 with the argument that these two antigens are used for the vaccines and therefore their antibodies should be included in the interpretation of positivity. The IgM Western blot is often positive in patients with persistent infection.6 Sometimes it is the only antibody marker detected.

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Columbia Lyme:

"Osp A 31 kd and Osp B 34 kd bands) are not even included in the top 10 CDC list but, if present with other specific bands, should be considered meaningful."

www.columbia-lyme.org/flatp/medworkup.html

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Journal of Immunology:

"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."

www.jimmunol.org/cgi/content/abstract/146/8/2776

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National Institutes of Health:

"Antibodies against a 35-kDa antigen of Borrelia burgdorferi are detectable in the serum of about half of patients with early Lyme disease.... Southern blotting revealed a chromosomal location for this gene; and it was specific for B. burgdorferi, B. afzellii, and B. garinii but not for B. hermsii, B. coriaciae, or B. turicatae."

www.pubmedcentral.nih.gov/articlerender.fcgi?artid=229516

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European Journal of Clinical Microbiology & Infectious Diseases:

"Abstract Sera of 52 Lyme borreliosis patients classified according to their clinical features were analysed by Western blot using as antigensBorrelia strains belonging to three recently described genomic species. The antibody response was demonstrated to be homologous within each genospecies. Serum reactivity was studied for each of the type strainsBorrelia burgdorferi sensu stricto (strain B31T),Borrelia garinii (strain 20047T) and group VS461. Seven of 15 sera (46.6 %) of patients with menin-goradiculitis showed preferential reactivity withBorrelia garinii (strain 20047T), all of 8 sera (100 %) of patients with acrodermatitis chronica atrophicans showed preferential reactivity with group VS461 (strain VS461) and 8 of 16 sera (50 %) of patients with arthritis showed preferential reactivity withBorrelia burgdorferi sensu stricto (strain B31T). The presence of a strong response to OspA and OspB proteins ofBorrelia burgdorferi sensu stricto was found only in this last group of patients. These results suggest that there are clinical implications of the recently described modifications in the taxonomy ofBorrelia burgdorferi."

www.springerlink.com/content/v2176801411q4512/

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British Journal of Biomedicine:

Some of the polypeptides from a B. burgdorferi extract may react with patient antibodies that are not specific to B. burgdorferi.4 These include the 41 kDa flagellin polypeptide, which, although it must be detected for a positive diagnosis, is known to cross-react with antibodies to proteins from other bacteria.4,5 Other polypeptides in this category include those of 44, 56, 62, 64, 72 and 82 kDa. In contrast, bands with 18, 22, 32, 34, 39, 46, 58 and 92 kDa polypeptides are generally accepted as specific for B. burgdorferi.1,6

Three of six parvovirus B19 IgM-positive serum samples reacted with specific B. burgdorferi antigens and the 41-kDa antigen (Table 1). The antibody response to parvovirus B19 is directed mainly against the two structural proteins of the viral capsid (VPl [83 kDa] and VP2 [58 kDa]).7,8 Although the molecular weights of these polypeptides are different from the B. burgdorferi-specific bands detected here (Table 1), they may share sufficient homology at particular epitopes for parvovirus B19-specific antibody to bind with B. burgdorferi-specific polypeptides.

Five of six CMV-IgM positive sera produced equivocal or weak positive B. burgdorferi Western blot results (Table 1). The individual structural proteins of CMV recognised by sera from IgG- and IgM-positive patients are 155, 149, 82.5, 74.5, 67, 57, 55, 38.5 and 28 kDa polypeptides.9 The 38.5 and 57 kDa polypeptides are of similar size to the 39 and 58 kDa polypeptides of B. burgdorferi, but there is no information about their homology. Other shared epitopes are suggested as the CMV IgM-positive sera also detect 34 and 92 kDa polypeptides on the B. burgdorferi blot.

In one case (sample 11, Table 1) the result was a weak positive. Although concurrent B. burgdorferi infection cannot be ruled out in the patients with parvovirus B19 and CMV infection whose sera cross-reacted, the B. burgdorferi IgG Western blot results from available previous or follow-up sera (samples 5, 8, 9, 11 and 12, Table 1) suggest that none of the patients had current or previous B. burgdorferi infection.

findarticles.com/p/articles/mi_qa3874/is_200607/ai_n17183134

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