RESUMO
DNA sequences encoding full-length outer surface protein (Osp) A were amplified from four joint fluid samples over 4.5 months from a patient with chronic Lyme arthritis, with a variant from wild type only found in sample 3. Rather than a mutation in vivo, these findings suggested a mixed infection in which BORRELIA: containing the wild-type and mutant ospA were waxing and waning in the patient's joint. If so, we reasoned that the mutant should be present in the community. We therefore took the novel epitope resulting from the mutation, expressed as a fusion protein in Escherichia coli, and performed Western blots on 80 high-titred stored sera; however, all except that of our index patient were negative. We then collected 36 stored sera from patients with Lyme disease residing within 10 miles of where the index patient had lived. An additional two sera from this circumscribed area were positive (P = 0.038). These findings show that results from single samples can be misleading, and suggest that the OspAs expressed in force late in Lyme arthritis are the same ones introduced initially into the host. Moreover, they allow a speculative mechanism for disease persistence not previously considered, in which antigenically distinct B. burgdorferi variant proteins present themselves serially to the immune system.
Assuntos
Antígenos de Superfície/genética , Proteínas da Membrana Bacteriana Externa/genética , Grupo Borrelia Burgdorferi/genética , Lipoproteínas , Doença de Lyme/microbiologia , Vacinas Bacterianas , Doença Crônica , Connecticut/epidemiologia , Feminino , Variação Genética , Humanos , Doença de Lyme/epidemiologia , Epidemiologia Molecular , Reação em Cadeia da Polimerase , Fatores de TempoRESUMO
Regulation of interleukin-1 (IL-1) mediated biological responses is complicated by the multiple ligands and receptors of the IL-1 family. Most studies of IL-1 receptors have used human or rodent cells. Here, we report that the coding region of the bovine type 1 interleukin-1 receptor (type 1 IL-1R) cDNA extends 1719 bp in length. Northern analysis of specific bovine cell and tissue RNA demonstrated a 4.5 kb transcript. Overall, the bovine type 1 IL-1R coding region exhibits approximately 81 and 76% similarity with the human type 1 IL-1R at the nucleotide and amino acid level, respectively, and somewhat less similarity with the mouse and rat sequences. Type 1 IL-1R transcripts were confirmed by RT-PCR in several bovine cell types, including peripheral blood mononuclear cells (PBMCs), neutrophils (PMNs), and fibroblast, peritoneal macrophage, and arterial endothelial cell lines. It is expected that molecular clones for the bovine type 1 and 2 IL-1 receptors will provide us with the tools needed to decipher species-and cell-specific regulation of IL-1 action in the bovine.
Assuntos
Bovinos/imunologia , DNA Complementar/química , Receptores de Interleucina-1/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , Clonagem Molecular , Humanos , Camundongos , Dados de Sequência Molecular , Ratos , Receptores de Interleucina-1/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Especificidade da EspécieRESUMO
The temporal synthesis of the P21 protein of Borrelia burgdorferi and the development of the humoral response to this antigen was assessed in infected mice. p21 is a member of the ospE-F gene family and its protein, P21, has been shown to be expressed by B. burgdorferi within infected mice but not by spirochetes cultured in vitro. P21 was not detected on B. burgdorferi in unfed or engorged Ixodes dammini (also known as I. scapularis) ticks, further supporting the postulate that P21 synthesis is specific for the mammalian host. In B. burgdorferi-infected mice, ospE mRNA and OspE antibodies were observed at 7 d, whereas p21 mRNA and P21-specific antibodies were detected at 21-28 d, suggesting that p21 is expressed later than ospE. Moreover, ospA mRNA was not discernible until day 14, indicating that ospA, like p21, is not expressed in the early stages of tick-transmitted murine Lyme borreliosis. Because p21 is expressed during infection in mice, we assessed the human humoral response to P21. 28% (34 of 122) of the patients with either early- or late-stage Lyme disease, and 33% (11 of 33) of the individuals with Lyme arthritis had P21 antibodies, suggesting that a P21 response may serve, at least partially, as a marker of infection. Active immunization with recombinant P21 did not protect C3H mice from tick-borne B. burgdorferi infection, and passive transfer of P21 antiserum to infected mice did not alter the course of disease. These data suggest that the antigenic structure of B. burgdorferi changes during the early stages of murine infection.