[Adverse unidentified transfusion reactions in five French regions: analysis, results, propositions]. / Les effets indésirables receveurs inconnus de cinq régions françaises: analyse, résultats, propositions.

OBJECTIVE: Adverse transfusion effects with "unidentified diagnosis" are yearly notified on the French national database "e-FIT" in various numbers and with high interregional discrepancies. The aim of this work was to analyse them in order do reach a better understanding of these notifications. RESULTS: On a total of 2499 "Fiches d'Effets Indésirables Receveurs" (FEIR) registered in 2006 in five French regions, 416 with "unidentified diagnosis" were analysed. Fifty-seven percent of them were kept classified in "unidentified diagnosis". Forty-three per cent of FEIR were reclassified, some in already proposed diagnostic categories (100 EIR), some in new proposed diagnostic categories (80 EIR) as: pathological context of the patient, pains linked to transfusion, hypotension. CONCLUSION: The study confirms the necessity to re-examine the French notification system. It underlines the insufficiency of clinical and biological investigations, which could allow to reach accurate diagnosis. It gives prominence the necessity of taking into account the patient's pathology and targets at least two diagnoses of transfusion adverse effects which are not yet proposed on the FEIR model. This work brings an overture about evolution of the French haemovigilance database which will involve further developments.

Bone marrow versus peripheral blood progenitor cells CD34 selection in patients with non-Hodgkin's lymphomas: different levels of tumor cell reduction. Implications for autografting.

Human CD34+ selected cells are able to reconstitute hematopoiesis in patients receiving a myeloablative treatment. Although the role of reinfused tumor cells contaminating the grafts on the determination of postautograft relapses remains unclear, the major interest of CD34+ cell selection is to reduce the tumor contamination of the graft. This can be achieved if tumor cells do not express the CD34 antigen. We previously showed that this approach was effective with bone marrow (BM) collections in patients with non-Hodgkin's lymphoma (NHL). Because peripheral blood progenitor cells (PBPC) allow faster hematologic recovery than BM and are expected to contain less tumor contamination, we have compared the results of CD34+ cell selection in 35 BM and 16 PBPC from 48 patients with NHL. The PBPC were collected after a course of chemotherapy followed by granulocyte colony-stimulating factor (G-CSF) administration. The data showed that the final CD34+ cell purity achieved with PBPC was higher than with BM (medians, 70% v 50%; P = .02). The CD34+ cell recovery was also better for PBPC (medians, 42% v 24%; P = .001). Tumor contamination was assessed by detection of BCL2/JH rearrangement using polymerase chain reaction (PCR) in 38 of 48 patients (22 BM, 16 PBPC). In addition, immunoglobulin heavy chain gene (IgH) rearrangements were investigated using PCR with consensus IgH primers. At harvesting, 10 of 22 BM and two of 16 PBPC contained BCL2/JH+ cells, one of 22 BM and 14 of 16 PBPC contained abnormal IgH+ cells (one PBPC contained both BCL2/JH+ and abnormal IgH+ cells) at harvesting. However, because lymphoma tissue specimens from patients at diagnosis were not available, the malignant character of IgH rearrangements could not be confirmed by sequencing and probing with allele-specific nucleotides. After CD34+ cell selection, a reduction to below the level of detection of BCL2/JH+ cells of BM and PBPC was effective in seven of 12 informative selections. In contrast, a reduction to below the level of detection of abnormal IgH+ cells was effective in only three of 15 informative selections. However, the detection of cells with an abnormal IgH pattern in the context of chemotherapy plus G-CSF progenitor mobilization in patients with NHL and its correlation with actual tumor contamination needs further investigation.

Human T-cell lymphotropic virus type I and II DNA amplification in seropositive and seronegative at-risk individuals.

To determine the presence or the absence of human T-lymphotropic virus type I and/or II (HTLV-I/II) DNA in at-risk individuals who were persistently negative for specific serologic assays, polymerase chain reaction with two primer pairs in common and conserved regions of HTLV-I and -II genomes was used. Seronegative individuals at risk for HTLV-I/II infection (15 heterosexual partners of seropositive individuals, 17 breastfed children born to HTLV-I-infected mothers, 47 multiply transfused patients, 22 intravenous drug users) were studied (n = 101); 35 seropositive individuals and 25 seronegative low-risk individuals were used as positive and negative controls, respectively. No positive polymerase chain reaction was observed in the seronegative at-risk individuals or in the negative controls. Positive controls gave positive results with at least one primer pair in all cases except one. A latent HTLV-I/II infection with a persistently negative serologic test for HTLV-I/II seems unlikely.

[Anti-HCV and recipients: situation in hemophiliacs in 1991]. / Anti-VHC et receveurs: situation chez les hémophiles en 1991.

Sera from 273 French haemophiliacs who had received non viral inactivated concentrates, were tested for antibodies to HCV by first and second generation assays. Antibodies to HCV were detected in 66% of the sera by the first generation assays (anti-C 100-3) reaching 100% by the second generation assays. None of the 53 patients only exposed to solvent-detergent treated Factor VIII or IX concentrates had HCV seroconversion. HCV core protein antibody was always detectable often as a single antibody in seropositive hemophilic patients.

No evidence of HIV-1 infection in seronegative hemophiliacs and in seronegative partners of seropositive hemophiliacs through polymerase chain reaction (PCR) and anti-NEF serology.

We prospectively studied a well-characterized cohort including 60 seronegative hemophiliacs or von Willebrand's disease patients, 6 seronegative female sexual partners of seropositive hemophiliacs, 59 seropositive hemophiliacs or von Willebrand's disease patients and 2 seropositive partners of seropositive hemophiliacs (used as positive controls), and 117 seronegative low risk individuals (used as negative controls). PCR assay, performed in peripheral blood mononuclear cells using three primer pairs in the gag, pol, LTR regions, showed no positive results in the 60 seronegative patients, in the 6 seronegative partners of seropositive patients and in the 117 seronegative low risk individuals, while PCR was positive with at least one primer pair in 53 (87%) of 61 seropositive patients. Anti-nef serology (Western-blot) was negative in seronegative patients, in seronegative partners of seropositive patients and positive in 58% out of the seropositive individuals. These results strongly suggest an absence of HIV-1 infection in individuals with a lastingly negative HIV serology.

Antibody to hepatitis C (anti C 100-3) in French hemophiliacs.

486 patients followed in four French hemophilia centres were tested for antibody to hepatitis C virus C100-3 recombinant protein. On samples collected in 1989, the overall incidence of anti-C 100-3 positivity was 66%. None of the 27 patients only exposed to solvent-detergent treated factor VIII or IX concentrates had C 100-3 antibodies. There was no difference according to the type of hemophilia nor to its severity. Serological follow-up from 1985 to 1989 was carried out on 51 patients. Two third of the C-100 3 paradoxically seronegative patients in 1989 were in fact positive for these antibodies within the last 5 years. They have lost their HCV antibodies as seen with the presently available C 100-3 test. Actually, as the virus has been already present at least for the last 10 years, (on 51 samples collected in 1979, the incidence of C-100-3 positivity was 78%), all treated patients may well have been in contact with HCV.

DNA amplification of HIV-1 in seropositive individuals and in seronegative at-risk individuals.

We assessed the HIV-1 status of seropositive and seronegative at-risk individuals by the polymerase chain reaction. Fifty-four out of 55 HIV-1-seropositive samples scored positive. However, HIV-1 proviral DNA was not detected in 16 seronegative homosexuals, 20 seronegative polytransfused haemophiliacs and 20 seronegative thalassaemic children, 20 individuals with isolated and persistent anti-core antibodies and 74 seronegative blood donors. These data indicate that positive HIV-1 DNA is likely to be an exceptional phenomenon in HIV-seronegative people.

[DNA amplification using the polymerase chain reaction method applied to the detection of HIV-1 in seropositive subjects and in seronegative subjects at risk]. / Amplification de l'ADN par la méthode de "Polymerase Chain Reaction" (PCR) appliquée à la détection de séquences VIH-1 chez des sujets séropositifs et des sujets séronégatifs à risque.

Polymerase Chain Reaction (PCR) allows the amplification of specific segments of Human Immunodeficiency Virus (HIV) genome. This technique is of interest for direct detection of HIV-1 DNA sequences in peripheral blood mononuclear cells. In our study, 54 out of 55 HIV-1 seropositive subjects were found positive with PCR assay. No detection of HIV DNA was observed in 36 seronegative at risk subjects (16 homosexuals and 20 polytransfused haemophiliacs), in 20 subjects with isolated and persistent anti-core antibodies, in 20 thalassemic polytransfused children and in 74 HIV seronegative blood donors (negative controls). These results are consistent with those of classical HIV serology and indicate that latent HIV-1 infection in seronegative at risk subjects is not a frequent event, if it exists.