Prevalence of IgG antibodies against Borrelia Burgdorferi s.l. and Ehrlichia Phagocytophila in sera of patients presenting symptoms of Lyme disease in a central region of Italy.

The aim of this study was to evaluate the prevalence (seroprevalence) of antibodies against Borrelia burgdorferi and Ehrlichia phagocytophila among patients resident in Lazio, a region of central Italy. Of a sample of 1,050 patients, which presented clinical manifestations related to Lyme disease, 34 (3.2%) were Borrelia-seropositive (Lyme index value ≥ 1.2). The sera of 25 out of the 34 patients that were Borrelia-positive were also analysed for the presence of antibodies against E. phagocytophila and 3 (12%) were found Ehrlichia-positive (titres >1:64). No Ehrlichia-positive samples were found among sera of 250 Borrelia-negative patients. Since both B. burgdorferi s.l. and Ehrlichia species share the same tick vector ( Ixodes ricinus), our results indicate that concurrent transmission of these microbial pathogens might have been occurred among the patients included in this study

Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells

<p>Abstract</p> <p>Background</p> <p>There is much evidence that tumor cells elicit a humoral immune response in patients. In most cases, the presence of antibodies in peripheral blood is detected only in small proportion of patients with tumors overexpressing the corresponding antigen. In the present study, we analyzed the significance of local humoral response provided by tumor-infiltrating lymphocytes in breast cancer patients.</p> <p>Methods</p> <p>The ability of a patient's immune system to produce specific antibodies inside tumor tissue, capable of recognizing tumor cells, was explored through analysis of the oligoclonality of antibodies derived from tumor-infiltrating lymphocytes and construction of a series of recombinant antibody libraries in scFv format, derived from breast tumor-infiltrating B lymphocytes. These libraries and one from peripheral blood lymphocytes of a single breast cancer patient were panned against three purified surface tumor antigens, such as CEA, MUC1 and ED-B domain, and against intact MCF7 breast carcinoma cells.</p> <p>Results</p> <p>Application of novel display vector, pKM19, allowed isolation of a large panel of breast cancer-specific antibodies against known tumor antigens, as well as against breast carcinoma cells. Reactivity of novel scFvs was confirmed by ELISA, immunohistochemistry, fluorescence staining and flow cytometry. We demonstrated that seven of ten primary breast tumor specimens, obtained using discarded surgical material, could be exploited as an appropriate source for generation of phage display libraries, giving highly specific antitumor antibodies which recognize heterologous tumor cells.</p> <p>Conclusion</p> <p>Local humoral immune response within tumor tissue in breast cancer patients frequently has an oligoclonal character. Efficient selection of specific antitumor antibodies from recombinant antibody libraries, derived from such oligoclonal tumor-infiltrated B lymphocytes, indicates the presence of natural immune response against tumor antigens in these patients. The described method is very promising for development of antitumor antibodies, potentially useful for diagnostic and therapeutic approaches.</p

Burkholderia cenocepacia Vaginal Infection in Patient with Smoldering Myeloma and Chronic Hepatitis C

We report a case of a vaginal infection caused by a strain of Burkholderia cenocepacia. The strain was isolated from vaginal swab specimens from a 68-year-old woman with smoldering myeloma and chronic hepatitis C virus infection who was hospitalized for abdominal abscess. Treatment with piperacillin/tazobactam eliminated B. cenocepacia infection and vaginal symptoms

Molecular characterization of Burkholderia cepacia isolates from cystic fibrosis (CF) patients in an Italian CF center

Bacteria of the Burkholderia cepacia complex consist of a number of closely related genomic species (genomovars) potentially pathogenic for cystic fibrosis (CF) patients, collectively referred to as the B. cepacia complex. The genomovar status and epidemiological relatedness of B. cepacia complex strains recovered from CF patients, attending a CF Center at the University Hospital “Policlinico Umberto I” of Rome, were investigated using 16S rRNA PCR-RFLP, recA PCR-RFLP, genomovar-specific PCR, and RAPD. Forty-seven isolates identified as B. cepacia by commercial systems were repeatedly recovered from 19 CF patients. The taxonomy approach used in this study showed that 17 of the 19 patients were colonized by B. cepacia complex strains. Genomovar III (11 strains) was the most prevalent genomovar. Two strains of genomovar I, one B. stabilis (genomovar IV), one B. multivorans (genomovar II), and 4 strains of B. anthina (genomovar VIII) were also identified. This is the first report of multiple patient colonization by B. anthina in a CF center. The epidemiological and genetic relatedness as well as the presence of molecular markers associated with virulence and transmissibility of the B. cepacia complex strains were determined and probable patient-to-patient spread was observed

Preclinical pharmacology and safety of a novel avidin derivative for tissue-targeted delivery of radiolabelled biotin

We recently described an oxidized avidin variant, named AvidinOX ®, which is a product that chemically links to tissue proteins while maintaining the capacity to uptake intravenously administered biotin. Such product proved to be successful in targeting radionuclide therapy in a mouse model of inoperable breast cancer. Here, we show that the uptake of a single or multiple doses of biotin (up to five times), by the tissue-bound AvidinOX ®, is stable for 2weeks. Taking into account that oxidized avidin is the first chemically reactive protein to be proposed for clinical use, we evaluated its tolerability, immunogenicity and mutagenicity. Present in vitro data indicate that AvidinOX ® (up to 10μg/5Ã\u9710 5 cells) does not affect cell viability or proliferation of PC3 human prostate cancer or 3T3 mouse fibroblast cell lines as well as primary mouse spleen cells. Safety pharmacology and toxicology studies were conducted using AvidinOX ® up to the highest concentration compatible with its solubility (about 12mg/mL), representing four times the product concentration intended for human use, and in the maximum administrable volume compatible with each study system. The intramuscular administration in rat and monkey induced a moderate to strong inflammatory response particularly after a second administration and consistently with the induction of an immune response. Interestingly, the intramuscular administration of AvidinOX ® to rodents and monkeys exhibiting very high anti-avidin antibody titres was well tolerated with no systemic symptoms of any kind. Intravenous administration of AvidinOX ®, performed to mimic an accidental injection of the dose intended for a local administration (15μL of 3.3mg/mL solution), showed significant localization of the product into the spleen not associated with uptake of the radiolabelled biotin intravenously injected after 24hr, thus suggesting rapid inactivation. No mutagenic activity was induced by oxidized avidin in prokaryotic and eukaryotic cells. Overall, the present data indicate that AvidinOX ® is well tolerated in rodents and non-human primates, thus supporting its clinical use within protocols of radionuclide therapy of inoperable tumour lesions. © 2011 The Authors. Basic & Clinical Pharmacology & Toxicology © 2011 Nordic Pharmacological Society

Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells-3

<p><b>Copyright information:</b></p><p>Taken from "Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells"</p><p>http://www.biomedcentral.com/1472-6750/7/70</p><p>BMC Biotechnology 2007;7():70-70.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2175506.</p><p></p>y genes. Corresponding PEG-purified phage was used as positive control (). The irrelevant anti-SP2 antibody gene of known origin, selected earlier from scFvEC23 library, was also tested

Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells-8

<p><b>Copyright information:</b></p><p>Taken from "Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells"</p><p>http://www.biomedcentral.com/1472-6750/7/70</p><p>BMC Biotechnology 2007;7():70-70.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2175506.</p><p></p>5, B87, B89, B90, B91, B92, B93, B95, B96), normal breast, normal testis and peripheral blood lymphocytes from four healthy donors (L1, L2, L3, L4), was used, as template for amplification of V(D)J antibody regions. Samples of cDNA were normalized by amplification of β-actin housekeeping gene. All V(D)J fragments were well-amplified and gave DNA bands of expected molecular weight in all cases, excluding normal testis cDNA sample. The same PCR products were fractionated by 10% PAGE, giving a higher resolution of DNA bands. Antibody subclass distribution. PCR-amplified normal breast and B84 cDNA samples not showing oligoclonal bands in V(D)J test, have prevalence of IgA bands in comparison to IgG1 and IgG2 (), while three samples, B91, B92 and B93, giving strong oligoclonal bands in previous test, have IgG1 or both IgG1 and IgG2 band prevalence in comparison with IgA (). Clonality of heavy chain antibodies derived from B92 and B93 cDNA samples. Amino acid sequences of variable regions of 30 clones were deduced from randomly sequenced γ-chain antibody genes derived from B92 and B93 cDNA. Peptide sequences are reported in single-letter code. Identical amino acids in similar clones are represented by a dash

Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells-7

<p><b>Copyright information:</b></p><p>Taken from "Tumor-infiltrating B lymphocytes as an efficient source of highly specific immunoglobulins recognizing tumor cells"</p><p>http://www.biomedcentral.com/1472-6750/7/70</p><p>BMC Biotechnology 2007;7():70-70.</p><p>Published online 18 Oct 2007</p><p>PMCID:PMC2175506.</p><p></p>gative control MCF10-2A cells was observed (data not shown)