LD1: a CD4−CD8− TCRαβ/CD3 + peripheral T cell line with helper function for B lymphocytes

In order to learn more about the small subset of CD4-CD8- TCRαβ/CD3 + peripheral T lymphocytes, we firstly characterized at the cellular and molecular levels the CD4-CD8- LD1 cell line isolated from the spleen of an MRL/lpr-lpr mouse. Secondly we studied its functional properties. LD1 cells are Thy1+ CD5 + CD4-CD8- LFA-1+ PgP-1+ and do not bind the T cell precursor-specific antibodies Joro 37.5 or Joro 75. They are negative for IgM, B-220, BP-1, J11d, Lyb8, la, F4/80, BP-2, and Mac-1 surface markers. LD1 cells have deleted the TCR5 locus, have rearrangements at the TCRγ gene cluster (i.e. a Vγ1 - Jγ1 -Cγ1 and a Vγ4.3-Jγ4-Cγ4) and have two rearrangements of the TCRβS gene cluster (I.e. a Dβ -Jβ1 and V-D-Jβ2). LD1 cells produce normal sized RNA transcripts from TCRα and β genes and lower levels of γ-mRNA. These cells bind CD3- and pan-TCRβ3-specific antibodies as determined by FM analysis. We conclude that LD1 cells bear a TCRαβ/CD3 type of receptor complex. LD1 cells fail both In vivo and in vitro to differentiate Into CD4+ or CD8 + cells. These cells help B lymphocytes to mature Into antibody-secreting cells, secrete IL-3 and IL-6 but not IL-2, IL-4, or IL-5, and exert no detectable cytolytlc activity. These results together with recent reports of antigen-specific CD4-CD8-TCRαβ/CD3+ cytotoxic T cell lines show that the CD4-CD8- TCRαβ/CD3+ subset comprises functionally competent helper and cytotoxic T lymphocytes and thereby argue for their potential to participate In immune responses. Our results also suggest that cells like LD1 represent terminally differentiated T lymphocytes rather than cells with precursor potential for CD4+ or CD8+ TCRαβ/CD3+ T lymphocyte

The combination of TRAIL and MG-132 induces apoptosis in both TRAIL-sensitive and TRAIL-resistant human follicular lymphoma cells

We have previously shown that the human follicular lymphoma cell line, HF28GFP, is sensitive to TRAIL-mediated apoptosis. Nevertheless, when the same cells overexpress anti-apoptotic Bcl-2 family protein, Bcl-xL (HF28Bcl-xL), they become resistant to TRAIL. Thus, these cell lines help us to investigate the action of novel apoptosis inducing candidate drugs. In the present study, we examined the effects of MG-132 (a proteasome inhibitor), LiCl (a glycogen synthase kinase-3 inhibitor) and/or TRAIL on pro-apoptotic Bcl-2 family proteins such as Bim and Bid. Here we demonstrate that the combination of MG-132 and TRAIL induced significant apoptotic cell death in both cell lines, HF28GFP and HF28BclxL. Apoptosis correlated with a decrease of phospho-ERK1/2, the accumulation of Bim and translocation of truncated Bid (tBid) and jBid. In addition, the combination of MG-132 and TRAIL seemed to target other apoptotic factors, which led to the accumulation of active capsase-3. Furthermore, co-stimulation of LiCl and TRAIL induced apoptosis in HF28GFP cells. However, HF28Bcl-xL cells were far less sensitive to the combinatorial effects of LiCl and TRAIL. Interestingly, we observed that LiCl did not target Bim and Bid proteins. In conclusion, these data show that targeting of pro-apoptotic Bcl-2 family proteins simultaneously through a selective proteasome inhibition might help to overcome TRAIL resistance caused by overexpression of anti-apoptotic Bcl-2 family proteins. Moreover, the data may provide new strategies to develop targeted therapies against lymphomas.Peer reviewe

RIP1 has a role in CD40-mediated apoptosis in human follicular lymphoma cells

CD40 is a cell surface receptor which belongs to tumor necrosis factor receptor (TNFR) family members. It transmits signals that regulate diverse cellular responses such as proliferation, differentiation, adhesion molecule expression and apoptosis. Unlike other TNFR family members (TRAIL-R, Fas-R and TNFR1), the CD40 cytoplasmic tail lacks death domain. However, CD40 is capable of inducing apoptosis in different types of cancer cells including lymphoma. The apoptotic effect of CD40 is linked to the involvement of Fas, TRAIL or receptor interacting protein 1 (RIP1) kinase. We have previously shown that CD40 activation has anti-apoptotic or apoptotic effect in follicular lymphoma (FL) cell lines. In this study, we investigated the mechanism by which CD40 mediates apoptosis in a follicular lymphoma cell line, HF4.9. We show here that CD40-induced apoptosis was dependent on caspase-8 activation because caspase-8 specific inhibitor, Z-IETD-FMK completely prevented apoptosis. Therefore, the involvement of TRAIL, Fas and RIP1 in caspase-8 activation was examined. The exogenous TRAIL-induced apoptosis was fully prevented by anti-TRAIL neutralizing antibody. However, the antibody had no effect on CD40-induced apoptosis indicating that CD40 did not induce the expression of endogenous TRAIL in HF4.9 cells. Moreover, the cells were not sensitive to Fas-mediated apoptosis. Interestingly, RIP1 specific inhibitor, necrostatin-1 decreased CD40-induced apoptosis, which showed that RIP1 has a role in caspase-8 activation. In conclusion, the survival or apoptotic effects of CD40-mediated signaling might be related to the differentiation stages of FL cells.Peer reviewe

Behaviour of Metals during Bioheap Leaching at the Talvivaara Mine, Finland

The behaviour of base metals Ni, Zn, Cu, Co, Fe, and Mn, potentially toxic metals Pb, Cr, and Cd, and the radioactive elements, U and Th, in the Talvivaara mining process, Finland has been studied by tracing metal concentrations from the black schist ore, through ores subjected to bioheap leaching of varying duration, to pregnant leach solution (PLS), and solid process waste material deposited on site in gypsum waste ponds. It is apparent that Zn, Cu, Co, and Cd are leached from the ore in a similar manner and recovered efficiently in the PLS; however, Ni, though leached, was also found in the gypsum pond at relatively high concentrations. Relatively little Pb is released from the ore, but the small fraction that is mobilised accumulates in the gypsum pond. Of the radioactive constituents, Th is essentially immobile, whereas U is readily leached from the ore, again accumulating in gypsum pond waste. In addition, a laboratory-based sequential leach test was applied to assess the future leaching potential of metals from residual ore and process waste material under different environmental conditions.Peer reviewe

Lapsille ja nuorille hyvä sote : LAPE-teesit sote-palveluiden uudistamisen tueksi

LAPE-teesit ovat suosituksia maakunnille ja yhteistyöalueille. Teeseissä kuvataan myös kuntien ja sivistystoimen näkökulmia tulevan maakunnan kanssa tehtävään yhteistyöhön. Teesit yhdistävät Lapsi- ja perhepalveluiden muutosohjelmassa (LAPE) työstetyt linjaukset, periaatteet ja uudet toimintamallit maakunta- ja sote-uudistuksen lainsäädäntökontekstiin (HE 15/2017 vp). LAPE-muutosohjelma perustuu lapsiin, nuoriin ja perheisiin liittyvään tieteelliseen tutkimukseen, vuosien aikana kertyneeseen kokemustietoon ja toimijoiden laajaan konsensukseen. Teesit tukevat maakunnissa ja kunnissa tehtävää pitkäjänteistä uudistamistyötä lasten, nuorten ja perheiden hyvinvoinnin varmistamiseksi myös vuoden 2018 jälkeen

Synergistic interaction in simultaneous exposure to Streptomyces californicus and Stachybotrys chartarum.

The microbial exposure associated with health complaints in moldy houses consists of a heterogeneous group of components, including both living and dead bacteria, fungi, and their metabolites and active compounds. However, little is known about the interactions between different microbes and their metabolites, although the cytotoxicity and inflammatory potential of certain individual microbes have been reported. In this study, we investigated the inflammatory responses of mouse RAW264.7 macrophages after exposure to six indoor air microbes (Aspergillus versicolor, Penicillium spinulosum, Stachybotrys chartarum, Bacillus cereus, Mycobacterium terrae, and Pseudomonas fluorescens) alone and together with the actinomycete Streptomyces californicus. The production of nitric oxide, levels of the proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6), and cytotoxicity were measured. The coexposure to Sta. chartarum and Str. californicus caused a synergistic increase in the production of IL-6 but not other cytokines. In further experiments, the metabolites from Sta. chartarum or from closely related fungi (atranones B and E, satratoxin G, trichodermin, 7-alpha-hydroxytrichodermol, staplabin, and SMTP-7) and the known fungal toxins sterigmatocystin, citrinin, and ochratoxin A were each tested with Str. californicus. The testing revealed a synergistic response in TNF-alpha and IL-6 production after coexposure to Str. californicus with both trichodermin and 7-alpha-hydroxytrichodermol. Finally, the synergistic inflammatory response caused by Str. californicus and trichodermin together was studied by analyzing for the presence of nuclear factor-kappa-B (NF-kappa-B) in nuclear extracts of the exposed cells. The exposure to Str. californicus induced the binding of NF-kappa-B proteins to the NF-kappa-B consensus sequence as well as to the natural NF-kappa-B site of the IL-6 promoter. Adding trichodermin to the exposure did not increase the DNA binding

Pleriksafori kantasolujen mobilisaatiossa autologista siirtoa varten

Vertaisarvioitu. English summaryAutologisella kantasolusiirrolla tuetulla intensiivihoidolla on keskeinen rooli etenkin multippelia myeloomaa sekä tietyissä tilanteissa lymfoomaa sairastavien potilaiden hoidossa. Huono tai epäonnistunut kantasolujen mobilisaatio ja keräys on tärkeä autologisen kantasolusiirron saatavuutta rajoittava tekijä. Noin vuosikymmenen ajan käytössä olleella pleriksaforilla voidaan usein auttaa potilaita, joiden kantasolut mobilisoituvat huonosti vaikuttamatta siirronjälkeiseen ennusteeseen. Pleriksaforin käytön on todettu vaikuttavan kerättyjen kantasolusiirteiden solukoostumukseen sekä potilaiden siirronjälkeiseen hematologiseen ja immunologiseen toipumiseen. Näiden löydösten kliininen merkitys vaatii vielä lisäselvityksiä, kuten optimaalisen autologisen kantasolusiirteen koostumuskin. Pleriksaforin optimaalisen ja etenkin kustannustehokkaan käytön kannalta on keskeistä tunnistaa oikeat potilasryhmät esimerkiksi tutkimuksissa varmennetuin algoritmein.Peer reviewe