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Identification of the immunological antigen-antibody reaction performed either in liquid phase or in agarose have several limits: antigen excess, low sensitivity, low speed reaction. A new method is described for the antigen-antibody reaction, easy, simple to perform and inexpensive. It is based on a reaction where the specific antibody is mixed with a semi-solid gel on a transparent surface and the antigen to be identified is inoculated into the gel. In this method, an immune precipitate can be shown in just few minutes, obtaining a good sensitivity compared with manual methods such as the Ouchterlony technique and other similar methods. By using albumin, IgG, RBP, transferrin, alpha-2-macroglobulin as antigen, the immune precipitate is formed within 10 min at 2–8 microgram/ml and, after 30 min, at 1–4 microgram/ml may be detected. The display of the immune precipitate is favored by oblique observation on a black background with a perpendicular light illuminating the gel from the bottom upwards. This method also allows a good control of antigen excess, a problem that is instead found in immunoturbidimetric reactions. The method here described has also been tested in serum and urine, and then it can be advantageously used as qualitative or semi-quantitative immunological test in research or diagnostic. Moreover, it could be particularly useful in low resources geographical areas, where an immunoassay that requires the use of dedicated equipment is not economically sustainable. The authors have waived the patent rights to allow the free development of new tests based on this method.

Identification of the immunological antigen-antibody reaction performed either in liquid phase or in agarose have several limits: antigen excess, low sensitivity, low speed reaction. A new method is described for the antigen-antibody reaction, easy, simple to perform and inexpensive. It is based on a reaction where the specific antibody is mixed with a semi-solid gel on a transparent surface and the antigen to be identified is inoculated into the gel. In this method, an immune precipitate can be shown in just few minutes, obtaining a good sensitivity compared with manual methods such as the Ouchterlony technique and other similar methods. By using albumin, IgG, RBP, transferrin, alpha-2-macroglobulin as antigen, the immune precipitate is formed within 10 min at 2–8 microgram/ml and, after 30 min, at 1–4 microgram/ml may be detected. The display of the immune precipitate is favored by oblique observation on a black background with a perpendicular light illuminating the gel from the bottom upwards. This method also allows a good control of antigen excess, a problem that is instead found in immunoturbidimetric reactions. The method here described has also been tested in serum and urine, and then it can be advantageously used as qualitative or semi-quantitative immunological test in research or diagnostic. Moreover, it could be particularly useful in low resources geographical areas, where an immunoassay that requires the use of dedicated equipment is not economically sustainable. The authors have waived the patent rights to allow the free development of new tests based on this method.

Novel agents in combination with melphalan and prednisone (MP) significantly improved progression-free survival (PFS) and overall survival (OS) in multiple myeloma (MM). Randomized trials comparing MP plus bortezomib (VMP) versus MP plus thalidomide (MPT) are lacking. Nine hundred and fifty-six elderly (>65 years) newly diagnosed MM patients from six European randomized trials were retrospectively analyzed and matched for age, albumin, and beta2-microglobulin at diagnosis, 296 patients were selected from the VMP groups, and 294 from MPT. Complete response rate was 21% in the VMP patients and 13% in the MPT patients (P = 0.007). After a median follow-up of 34 months (range, 1-92), VMP significantly prolonged both PFS (median 32.5 vs. 22.9 months, HR 0.65; 95% CI 0.52-0.82; P < 0.001) and OS (median 79.7 vs. 45.1 months, HR 0.44; 95% CI 0.32-0.59; P < 0.001) in comparison with MPT. The benefit in terms of OS of the VMP group was quite similar among patients with different risk factors defined by sex, ISS, ECOG performance status, or serum creatinine but not among patients ≥ 75 years. Multivariate analysis confirmed that VMP was an independent predictor of longer PFS and OS. In a control-case matched analysis, PFS and OS were prolonged in patients who received VMP in comparison with those treated with MPT.

Novel agents in combination with melphalan and prednisone (MP) significantly improved progression-free survival (PFS) and overall survival (OS) in multiple myeloma (MM). Randomized trials comparing MP plus bortezomib (VMP) versus MP plus thalidomide (MPT) are lacking. Nine hundred and fifty-six elderly (>65 years) newly diagnosed MM patients from six European randomized trials were retrospectively analyzed and matched for age, albumin, and beta2-microglobulin at diagnosis, 296 patients were selected from the VMP groups, and 294 from MPT. Complete response rate was 21% in the VMP patients and 13% in the MPT patients (P = 0.007). After a median follow-up of 34 months (range, 1-92), VMP significantly prolonged both PFS (median 32.5 vs. 22.9 months, HR 0.65; 95% CI 0.52-0.82; P < 0.001) and OS (median 79.7 vs. 45.1 months, HR 0.44; 95% CI 0.32-0.59; P < 0.001) in comparison with MPT. The benefit in terms of OS of the VMP group was quite similar among patients with different risk factors defined by sex, ISS, ECOG performance status, or serum creatinine but not among patients ≥ 75 years. Multivariate analysis confirmed that VMP was an independent predictor of longer PFS and OS. In a control-case matched analysis, PFS and OS were prolonged in patients who received VMP in comparison with those treated with MPT.

Non-myeloablative stem cell transplantation (NMT) has been increasingly used in compromised patients who would otherwise have been unable to undergo allotransplant. There is little understanding of the kinetics of immune reconstitution and its influence on infective complications following NMT. The aim of present study was to evaluate lymphocyte subset reconstitution over the first 12 months post-transplant in 15 adult patients receiving NMT with comparison to that of 30 patients grafted with a conventional hemopoietic stem cell transplantation (HSCT). NMT recipients were conditioned with fludarabine-based conditioning regimens. Peripheral blood stem cell (PBSC) was the source of stem cells in 13 NMT recipients and in 24 conventional HSCT recipients. Absolute numbers of helper (CD4+) T cells, naive (CD4+ CD45RA+) and memory (CD4+ CD45RO+) T cells as well as suppressor (CD8+) T cells, CD19+ B cells and NK cells were comparable in the two groups at all time points after transplantation. A median value of 200 CD4+ T cells/microl was achieved at 2 months post-transplant by the NMT and HSCT recipients. The CD4:CD8 ratio remained severely depressed throughout the study period. Almost all CD4+ lymphocytes expressed CD45RO antigen in the both groups of patients B lymphocytes showed low counts throughout the entire study period in both groups. Bacteremia and CMV antigenemia occurred respectively in 13 and 36% of the patients in the NMT group and in 15 and 39% of the patients in the HSCT group. Our preliminary data indicate that patients receiving a NMT have a lymphocyte reconstitution similar to that observed in patients who received a conventional HSCT. The incidence of bacteremia and CMV infection were not significantly different between the groups. Nevertheless, due to the small sample size, these results should be considered suggestive rather than definitive.