MgpB Types among Mycoplasma genitalium Strains from Men Who Have Sex with Men in Berlin, Germany, 2016-2018.

Mycoplasma genitalium is a cell wall-less bacterium causing urethritis and other sexually transmitted diseases. Despite a strongly conserved genome, strains in clinical samples can be typed by different methods. To obtain data from the risk population of men having sex with men, we analyzed the typing region in the gene coding for the MgpB adhesin of M. genitalium first in 163 and 45 follow-up samples among patients of two specialized practices in Berlin, Germany. Strains belong to 43 different mgpB types emphasizing the diversity of the genome region. With respect to 133 types previously described, 27 new types were found. However, the majority of strains (64.4%) were assigned to types 4, 6, 113, and 108, respectively. A correlation between mgpB type and the occurrence of mutations associated with macrolide and quinolone resistance was not demonstrated. Investigation of follow-up samples from 35 patients confirmed the same mgpB and, additionally, MG_309 types in 25 cases. In 10 cases, differences between types in subsequent samples indicated an infection with a genetically different strain in the period between samplings. MgpB/MG_309 typing is a useful method to compare M. genitalium strains in samples of individual patients as well as those circulating in different populations.

Prevalence of macrolide- and fluoroquinolone-resistant Mycoplasma genitalium strains in clinical specimens from men who have sex with men of two sexually transmitted infection practices in Berlin, Germany.

OBJECTIVES: The cell-wall-less Mollicutes species Mycoplasma genitalium is a sexually transmitted micro-organism that causes different male and female genital tract infections. In recent years, resistance of the pathogen to macrolides and fluoroquinolones has been increasingly reported worldwide and is more frequent in risk groups. METHODS: To determine the rates of antimicrobial resistance, M. genitalium strains in 195 specimens from 154 outpatients (154 first and 41 follow-up samples) treated in two specialised practices between September 2017 and December 2018 in Berlin, Germany, were analysed. RESULTS: The included patients were predominantly men who have sex with men (MSM) (91.6%) and were HIV-positive in many cases (49.4%). Only 27.3% of M. genitalium-positive patients reported symptoms. Among the first samples (mainly rectal swabs) (57.8%), mutations associated with macrolide (23S rRNA) and quinolone (parC gene) resistance were detected in 79.9% and 13.0% of strains, respectively. Resistance to both classes of antibiotics was found in 11.7% of specimens. Changes of A→G at position 2072 of 23S rRNA and of serine at position 83 of ParC were the most frequent alterations. CONCLUSION: Although azithromycin is recommended as a first-line antibiotic to treat infections with M. genitalium in MSM, according to these data its use must be highly limited in Berlin. Besides the need for resistance studies regarding strains circulating in other locations and among different patient groups in Germany, the results emphasise the importance of intensified antibiotic resistance testing of M. genitalium to avoid a further increase in treatment failures in infections with this emerging human pathogen.

Transfer of Autologous Gene-modified T Cells in HIV-infected Patients with Advanced Immunodeficiency and Drug-resistant Virus.

Drug toxicity and viral resistance limit the long-term efficacy of antiviral drug treatment for human immunodeficiency virus (HIV) infection. Thus, alternative therapies need to be explored. We tested the infusion of T lymphocytes transduced with a retroviral vector (M87o) that expresses an HIV entry-inhibitory peptide (maC46). Gene-modified autologous T cells were infused into ten HIV-infected patients with advanced disease and multidrug-resistant virus during anti-retroviral combination therapy. T-cell infusions were tolerated well, with no severe side effects. A significant increase of CD4 counts was observed after infusion. At the end of the 1-year follow-up, the CD4 counts of all patients were still around or above baseline. Gene-modified cells could be detected in peripheral blood, lymph nodes, and bone marrow throughout the 1-year follow-up, and marking levels correlated with the cell dose. No significant changes of viral load were observed during the first 4 months. Four of the seven patients who changed their antiviral drug regimen thereafter responded with a significant decline in plasma viral load. In conclusion, the transfer of gene-modified cells was safe, led to sustained levels of gene marking, and may improve immune competence in HIV-infected patients with advanced disease and multidrug-resistant virus.

A phase 1 study to evaluate the safety and immunogenicity of a recombinant HIV type 1 subtype C adeno-associated virus vaccine.

A novel prophylactic AIDS vaccine candidate, consisting of single-stranded DNA for HIV-1 subtype C gag, protease, and part of reverse transcriptase genes, enclosed within a recombinant adeno-associated virus serotype-2 protein capsid (tgAAC09) induced T cell responses and antibodies in nonhuman primates. In this randomized, dose escalation phase I trial, HIV-uninfected healthy volunteers (50 in Europe, 30 in India) received a single intramuscular injection of tgAAC09 at 3 x 10(9) DNase resistant particles (DRP) (n = 16), 3 x 10(10) DRP (n = 23), 3 x 10(11) DRP (n = 25), or placebo (n = 16). Twenty-one participants in Europe received a second (boost) dose of 3 x 10(11) DRP tgAAC09 or placebo at least 24 weeks after the first injection. The vaccine was safe and well-tolerated after initial and boost vaccination. Local and systemic reactogenicity was experienced by 13-25% of participants and was not dose related. No vaccine-related serious adverse events were reported. Modest HIV-specific T cell responses were detected in 7/64 vaccinees (40-385 SFC/10(6) PBMC), with 16% (4/25) responders in the highest dose group. All responses were to Gag epitopes. tgAAC09 appears to be safe, well-tolerated, and modestly immunogenic. Further evaluation of higher doses of tgAAC09 and boost injections is ongoing in Africa.

Transfer of autologous gene-modified T cells in HIV-infected patients with advanced immunodeficiency and drug-resistant virus.

Drug toxicity and viral resistance limit the long-term efficacy of antiviral drug treatment for human immunodeficiency virus (HIV) infection. Thus, alternative therapies need to be explored. We tested the infusion of T lymphocytes transduced with a retroviral vector (M87o) that expresses an HIV entry-inhibitory peptide (maC46). Gene-modified autologous T cells were infused into ten HIV-infected patients with advanced disease and multidrug-resistant virus during anti-retroviral combination therapy. T-cell infusions were tolerated well, with no severe side effects. A significant increase of CD4 counts was observed after infusion. At the end of the 1-year follow-up, the CD4 counts of all patients were still around or above baseline. Gene-modified cells could be detected in peripheral blood, lymph nodes, and bone marrow throughout the 1-year follow-up, and marking levels correlated with the cell dose. No significant changes of viral load were observed during the first 4 months. Four of the seven patients who changed their antiviral drug regimen thereafter responded with a significant decline in plasma viral load. In conclusion, the transfer of gene-modified cells was safe, led to sustained levels of gene marking, and may improve immune competence in HIV-infected patients with advanced disease and multidrug-resistant virus.