[Impact of anti-HLA antibodies on outcomes of allogeneic stem cell transplantation: a report by the SFGM-TC]. / Impact des anticorps anti-HLA sur le devenir de l'allogreffe de cellules souches hématopoïétiques : un rapport par la SFGM-TC.

The role of anti-HLA antibodies in allogeneic stem cell transplantation setting is still unclear. In the attempt to harmonize clinical practices between different French transplantation centers, the French Society of Bone Marrow Transplantation and Cell Therapies (SFGM-TC) set up its fourth annual series of workshops which brought together practitioners from all of its member centers. These workshops took place in September 2013 in Lille. This article offers the recommendations of the group that considered the impact that have anti-HLA antibodies on outcomes in allogeneic stem cell transplantation.

Transfusion-related acute lung injury after intravenous immunoglobulin treatment in a lung transplant recipient.

Three weeks after single-lung transplantation for pulmonary fibrosis, a patient with high serum levels of de novo donor-specific antibodies received high-dose intravenous immunoglobulin (IVIG) infusion (scheduled dose: 2 g/kg on 2 days) to prevent antibody-mediated rejection. Within the first hours after completion of infusions, he experienced acute lung injury involving the transplanted lung. Given the clinical evolution and the absence of an alternative diagnosis, transfusion-related acute lung injury (TRALI) was diagnosed. The IVIG administered on each day was from the same batch. At day 110, because of an increase in the serum titers of donor-specific antibodies, IVIG therapy was reintroduced but from a different batch, with excellent clinical tolerance. The lung injury was explored biologically, but no mechanism was revealed. Given the increasing use of IVIG in solid-organ recipients, clinicians should be aware of possible TRALI after IVIG infusion.

[Caracterization of HLA allo-immunization and clinical impact in transfusion and organ transplantation]. / Caractérisation de l'allo-immunisation anti-HLA et impact clinique en transfusion et en transplantation d'organe.

Allo-immunizations against HLA antigens are known to be deleterious in transfusion and organ transplantation. The development of new tests based on solid phase assays for screening and identification of HLA antibodies in particular those using Luminex® bead based technology has completely changed the way of allo-immunization monitoring because of their extreme sensitivity. They allow a better characterization of these antibodies, identification of acceptable antigens and the use of virtual cross-matches. All these new possibilities improve the managing of patients before and after platelets transfusion or organ transplantation. However, this technology displays some limits that should be known in order to interpret correctly the results. Beside these bead based assays, cellular cross-matches based on Complement Dependent Cytotoxicity (CDC) and flow cytometry are still used and useful in organ transplantation since beads are produced in vitro and do not reflected exactly what happens physiologically. Moreover, differences of sensitivity between these methods make results interpretation and decision making difficult in some cases.

Two novel human and mouse DNA polymerases of the polX family.

We describe here two novel mouse and human DNA polymerases: one (pol lambda) has homology with DNA polymerase beta while the other one (pol mu) is closer to terminal deoxynucleotidyltransferase. However both have DNA polymerase activity in vitro and share similar structural organization, including a BRCT domain, helix-loop-helix DNA-binding motifs and polymerase X domain. mRNA expression of pol lambda is highest in testis and fetal liver, while expression of pol mu is more lymphoid, with highest expression both in thymus and tonsillar B cells. An unusually large number of splice variants is observed for the pol mu gene, most of which affect the polymerase domain. Expression of mRNA of both polymerases is down-regulated upon treatment by DNA damaging agents (UV light, gamma-rays or H(2)O(2)). This suggests that their biological function may differ from DNA translesion synthesis, for which several DNA polymerase activities have been recently described. Possible functions are discussed.

A novel allele HLA-C*07:445 identified in a French hematopoietic stem cell donor.

The novel allele HLA-C*07:445 has 1 nucleotide change from HLA-C*07:01 at nucleotide 277 C>A in exon 2.

Risk factors and outcome of graft failure after HLA matched and mismatched unrelated donor hematopoietic stem cell transplantation: a study on behalf of SFGM-TC and SFHI.

Graft failure remains a severe complication of hematopoietic stem cell transplantation (HSCT). Several risk factors have already been published. In this study, we re-evaluated them in a large cohort who had the benefit of the recent experience in HSCT (2006-2012). Data from 4684 unrelated donor HSCT from 2006 to 2012 were retrospectively collected from centers belonging to the French Society for Stem Cell Transplantation. Among the 2716 patients for whom HLA typing was available, 103 did not engraft leading to a low rate of no engraftment at 3.8%. In univariate analysis, only type of disease and status of disease at transplant for malignant diseases remained significant risk factors (P=0.04 and P<0.0001, respectively). In multivariate analysis, only status of disease was a significant risk factor (P<0.0001). Among the 61 patients who did not engraft and who were mismatched for 1 HLA class I and/or HLA-DP, 5 donor-specific antibodies (DSAs) were detected but only 1 was clearly involved in graft failure, for the others their role was more questionable. Second HSCT exhibited a protective although not statistically significant effect on OS (hazard ratio=0.57 [0.32-1.02]). In conclusion, only one parameter (disease status before graft) remains risk factor for graft failure in this recent cohort.

Is there any impact of HLA-DPB1 disparity in 10/10 HLA-matched unrelated hematopoietic SCT? Results of a French multicentric retrospective study.

We retrospectively analyzed the impact of HLA-DPB1 mismatches in a large cohort of 1342 French patients who underwent 10/10 HLA-matched unrelated HSCT. A significant impact of HLA-DPB1 allelic mismatches (2 vs 0) was observed in severe acute GVHD (aGVHDIII-IV) (risk ratio (RR)=1.73, confidence interval (CI) 95% 1.09-2.73, P=0.019) without impact on OS, TRM, relapse and chronic GVHD (cGVHD). According to the T-cell epitope 3 (TCE3)/TCE4 HLA-DPB1 disparity algorithm, 37.6% and 58.4% pairs had nonpermissive HLA-DPB1, respectively. TCE3 and TCE4 disparities had no statistical impact on OS, TRM, relapse, aGVHD and cGVHD. When TCE3/TCE4 disparities were analyzed in the graft-vs-host or host-vs-graft (HVG) direction, only a significant impact of TCE4 nonpermissive disparities in the HVG direction was observed on relapse (RR=1.34, CI 95% 1.00-1.80, P=0.048). In conclusion, this French retrospective study shows an adverse prognosis of HLA-DPB1 mismatches (2 vs 0) on severe aGVHD and of nonpermissive TCE4 HVG disparities on relapse after HLA-matched 10/10 unrelated HSCT.

Tn3702, a conjugative transposon in Enterococcus faecalis.

Enterococcus faecalis strain D434 was found to carry on its chromosome a determinant encoding tetracycline-minocycline resistance (Tcr-Mnr) and to harbor both an R plasmid and a cryptic conjugative plasmid, pIP1141. The determinant coding for Tcr-Mnr was located on a conjugative transposon, designated Tn3702. The transposition of Tn3702 on to both pIP1141 and the hemolysin plasmid pIP964 yielded different derivatives each of which contained an 18.5-kilobase insert. The structure of Tn3702 is similar to that of the conjugative transposon Tn916.

[Viridans streptococci in human infections: identification and susceptibility to antibiotics]. / Les streptocoques non groupables dans les infections humaines: identification et sensibilité aux antibiotiques.

A method for the speciation of viridans streptococci (devoided of group antigens) is described. The major identification criteria are based on the reaction of a series of biochemical tests such as acid production in lactose, inuline, raffinose, mannitol and sorbitol, hydrolysis of arginine, esculin and Na hippurate, and production of polysaccharides in 5% sucrose media. A total of 460 strains were isolated from human specimens and identified as follows: 118 Streptococcus mitis, 102 S. sanguis II, 75 S. Sanguis I, 87 S. milleri (Streptococcus MG-intermedius), 28 S. mutans, 25 S. salivarius, 14 S. morbillorium, 2 S. uberis and 9 unspeciated. Susceptibility to antibiotics was studied for 318 strains: 63% of them were susceptible to all drugs tested; 37% of the strains were resistant to one or several antibiotics as follows: 34% to tetracycline, 8.5% to macrolides and related drugs, 5.3% to streptomycin and/or kanamycin (MIC greater than 2,000 micrograms/ml), 5% to penicillin (MIC = 1-4 micrograms/ml) and 4% to chloramphenicol.

[Group D streptococci in human infections: identification and sensitivity to antibiotics (author's transl)]. / Les streptocoques du groupe D dans les infections humaines: identification et sensibilité aux antibiotiques.

A method for the speciation of group D streptococci is described. A major criterion for identification is the reaction of antigenic extracts against streptococcal group D antisera. In addition, a series of biochemical tests is used: bile-esculine, resistance to 6.5% sodium chloride and potassium tellurite, acid production in mannitol, sorbitol and raffinose, hydrolysis of starch and arginine, and production of dextran. A total of 184 strains was isolated from human material and identified as follows: 104 S. faecalis, 18 S. faecium, 8 S. durans, 2 S. avium, 51 s. bovis and 1 unspeciated. The sensitivity to antibiotics was studied for 141 strains: 34% were sensitive, 23% were singly resistant to tetracycline and 43% were multiply resistant (tetracycline, macrolides and related drugs, high-level resistance to aminoglycosides and to chloramphenicol).

Viridans streptococci in infective endocarditis: species distribution and susceptibility to antibiotics.

A method for the speciation of viridans streptococci (non-groupable) is described. The major identification criteria are based on the reactions to a series of biochemical tests, including acid production in lactose, inulin, raffinose, mannitol and sorbitol, hydrolysis of arginine, esculin and Na hippurate, and production of polysaccharides in 5% sucrose media. A total of 450 strains was isolated from blood cultures, 183 of which were from confirmed cases of subacute endocarditis. The latter were identified as follows (%): Streptococcus sanguis I (25.7), S. mitis (19.7), S. sanguis II (19.7), S. mutans (17.5), S. milleri (12), S. morbillorium (3.2) and S. salivarius (2.2). Susceptibility to antibiotics was studied for 129 of these strains: 68% were susceptible to all drugs tested, 20% were resistant only to tetracycline, 4% only to penicillin (MIC = 0.5-4 micrograms ml-1) and 8% were multiply resistant (tetracycline, macrolides and related drugs, chloramphenicol, penicillin, high-level resistance to kanamycin and/or streptomycin [MIC = 1000-80.00 micrograms ml-1].

Does a tetracycline resistance determinant of class N exist?

pMV120 was reported to carry the tetracycline resistance (Tcr) determinant of class N. We obtained tetracycline-susceptible transconjugants harboring plasmids with restriction enzyme profiles indistinguishable from those of pMV120 isolated from tetracycline-resistant clones. We conclude that pMV120 is a cryptic plasmid and that class N of Tcr determinants does not exist.

Diversity of chromosomal genetic elements and gene identification in antibiotic-resistant strains of Streptococcus pneumoniae and Streptococcus bovis.

Antibiotic-resistant Streptococcus pneumoniae (26 strains) and Streptococcus bovis (28 strains), devoid of R plasmids, were examined for DNA-DNA homology to Tn916 and Tn3701. Tn916-like structures were found in 17 S. pneumoniae and 21 S. bovis strains. Tn916-modified structures were present in 6 S. pneumoniae and 2 S. bovis strains. Two strains of each species carried elements having a Tn3701-like composite structure. All these elements were chromosome-borne. No chromosomal elements were detected in 1 S. pneumoniae and 3 S. bovis strains.

[Characteristics of "Streptococcus mutans" from endocarditis and susceptibility to antimicrobial agents (author's transl)]. / Caractéristiques des souches de Streptococcus mutans isolées d'endocardites subaigues et sensibilité aux antibiotiques.

Strains of Streptococcus mutans were isolated from blood cultures of ten patients with endocarditis. Nine of these patients had a typical clinical picture of subacute bacterial endocarditis, with fever, weakness, heart murmur and multiple positive blood cultures. All the patients had previous valvular heart diseases; only in three cases the initiating event involved some type of dental manipulations which where supposed as the source of infection. The major criteria for recognizing S. mutans were colony morphology on blood agar, characteristic extracellular polysaccharide production in 5% sucrose broth, acid formation in mannitol and sorbitol broth, and the failure of antigenic extracts of S. mutans to react with streptococcal group antisera. The susceptibility to antimicrobial agents was tested by the diffusimetric method with susceptibility disks. All the strains were susceptible to penicillin G, erythromycin, pristinamycin, lincomycin and tetracycline, and resistant to streptomycin and gentamicine.

High-level aminoglycoside resistance in group A, B, G, D (Streptococcus bovis), and viridans streptococci.

Of 20 clinical isolates of group A, B, G, D (Streptococcus bovis), and viridans streptococci, 5 transferred their antibiotic resistance markers into streptococcal recipients at a low frequency (10(-4) to 10(-8)) in the apparent absence of extrachromosomal elements. All strains carried genetic markers for high-level resistance to streptomycin, kanamycin, neomycin, lividomycin A, and ribostamycin, as well as resistance to macrolides and related drugs, tetracycline, and chloramphenicol.