Peripheral blood-derived, γ9δ2 t cell-enriched cell lines from glioblastoma multiforme patients exert anti-tumoral effects in vitro.

The goal of this work was to assess the potential of T cells expressing Vγ9Vδ2+ T cell receptors (TCR, γ9δ2T cells) present in peripheral blood (PB) m ononuclear cells (MC, PBMC) of glioblastoma multiforme (GBM) patients to act as anti-tumoral agents. We found that γ9δ2T cell levels were decreased in patients' PB relative to a cohort of healthy donors (HD) (respectively 0.52±0.55%, n=16, vs 1.12±0.6%, n=14, p=0.008) but did not significantly correlate with postoperative survival (R=0.6, p=0.063). Importantly, however, the γ9δ2T cells could be expanded in vitro to consist 51±23% of the cultured lymphocytes (98% CD3+). This was achieved after 14 days of culture in medium containing the amino-bisphosphonate (ABP) Zoledronate (Zol) and interleukin (IL)-2, resulting in γ9δ2T cell-enriched lines (gdTCEL) similar to those of HD derived gdTCEL (54±19%). Moreover, gdTCEL from patients and HD mediated cytotoxicity to GBM-derived cell lines (GBMDCL), which was abrogated by immune-magnetic removal of the γ9δ2T cells. Furthermore, low level interferon (IFN) γ secretion was induced by gdTCEL briefly co-cultured with GBMDCL or autologous - tumor-derived cells, which was greatly amplified in the presence of Zol. Importantly, IFNγ secretion was inhibited by mevastatin but enhanced by cross-linking of butyrophilin 3A1 (CD277) on a CD277+ GBMDCL (U251MG) or by pretreatment of GBMDCL with temozolomide (TMZ). Taken together, these data suggest that γ9δ2T cells in PB of GBM patients can give rise to gdTCEL that mediate anti-tumoral activities.

A certified plasmid reference material for the standardisation of BCR-ABL1 mRNA quantification by real-time quantitative PCR.

Serial quantification of BCR-ABL1 mRNA is an important therapeutic indicator in chronic myeloid leukaemia, but there is a substantial variation in results reported by different laboratories. To improve comparability, an internationally accepted plasmid certified reference material (CRM) was developed according to ISO Guide 34:2009. Fragments of BCR-ABL1 (e14a2 mRNA fusion), BCR and GUSB transcripts were amplified and cloned into pUC18 to yield plasmid pIRMM0099. Six different linearised plasmid solutions were produced with the following copy number concentrations, assigned by digital PCR, and expanded uncertainties: 1.08±0.13 × 10(6), 1.08±0.11 × 10(5), 1.03±0.10 × 10(4), 1.02±0.09 × 10(3), 1.04±0.10 × 10(2) and 10.0±1.5 copies/µl. The certification of the material for the number of specific DNA fragments per plasmid, copy number concentration of the plasmid solutions and the assessment of inter-unit heterogeneity and stability were performed according to ISO Guide 35:2006. Two suitability studies performed by 63 BCR-ABL1 testing laboratories demonstrated that this set of 6 plasmid CRMs can help to standardise a number of measured transcripts of e14a2 BCR-ABL1 and three control genes (ABL1, BCR and GUSB). The set of six plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (https://ec.europa.eu/jrc/en/reference-materials/catalogue/; CRM code ERM-AD623a-f).

Intraocular pressure control and fluctuation: the effect of treatment with selective laser trabeculoplasty.

AIMS: To evaluate the effect of selective laser trabeculoplasty (SLT) on intraocular pressure (IOP) control and diurnal tension curves of patients with open-angle glaucoma (OAG) and ocular hypertension (OHT), and to compare this effect with that of latanoprost. METHODS: Forty patients were randomised to receive either SLT or latanoprost. IOP control was evaluated by comparing pretreatment values with post-treatment measurements on day 3, week 1, month 1 and 4-6 months; success was defined as 20% decrease in IOP. Tension curves were plotted prior to treatment and 4-6 months afterwards; success was 50% reduction in fluctuation. RESULTS: SLT decreased pressure by 4.7 mm Hg on average (95% CI 3.6 to 5.7 mm Hg; p<0.01). The reduction was similar for latanoprost at all follow-ups except month 1; 75% of SLT patients and 73% of latanoprost patients achieved success in IOP control (p = 0.4). SLT significantly reduced IOP fluctuation, but latanoprost was more effective (3.6 mm Hg, 95% CI 3.2 to 3.9 mm Hg vs 2.5 mm Hg, 95% CI 2.2 to 2.9 mm Hg for SLT; p = 0.04). Success in fluctuation reduction was 50% for SLT and 83% for latanoprost (p = 0.045). CONCLUSIONS: Both SLT and latanoprost had a significant impact on IOP control and fluctuation. While latanoprost may be more likely to reduce IOP fluctuation, SLT has the benefit of being a one-time intervention not requiring ongoing patient compliance.

A randomised, prospective study comparing selective laser trabeculoplasty with latanoprost for the control of intraocular pressure in ocular hypertension and open angle glaucoma.

AIM: To compare 90 degrees , 180 degrees , and 360 degrees selective laser trabeculoplasty (SLT, 532 nm Nd:YAG laser) with latanoprost 0.005% for the control of intraocular pressure (IOP) in ocular hypertension (OHT) and open angle glaucoma (OAG). METHODS: A prospective, randomised clinical trial in the Department of Ophthalmology, St Thomas's Hospital, London, and Clayton Eye Centre, Wakefield, West Yorkshire. 167 patients (167 eyes) with either OHT or OAG were randomised to receive 90 degrees , 180 degrees , and 360 degrees SLT or latanoprost 0.005% at night and were evaluated at 1 hour, 1 day, 1 week and 1, 3, 6, and 12 months. RESULTS: The mean follow up was 10.3 months (range 1--12 months). Early, transient, complications such as postoperative ocular pain, uveitis, and 1 hour IOP spike occurred in a number of eyes after SLT, with pain being reported more frequently after 360 degrees than 90 degrees treatments (p>0.001). Success rates defined in terms of both a 20% or more and a 30% or more IOP reduction from baseline measurements with no additional antiglaucomatous interventions were better with latanoprost than 90 degrees (p<0.001) and 180 degrees SLT (p<0.02) treatments. Differences in success rates between latanoprost and 360 degrees SLT did not reach statistical significance (p<0.5). Success rates were greater with 180 degrees and 360 degrees compared to 90 degrees SLT (p<0.05). With 360 degrees SLT, 82% of eyes achieved a >20% IOP reduction and 59% a >30% reduction from baseline. Although success rates were better with 360 degrees than 180 degrees SLT treatments, differences did not reach statistical significance. There were no differences with regard to age, sex, race, pretreatment IOP, OHT versus OAG, laser power settings, and total laser energy delivered between eyes which responded, in terms of a >20% and a >30% IOP reduction, and those that did not respond with 180 degrees and 360 degrees SLT treatments. CONCLUSIONS: Success rates were higher with latanoprost 0.005% at night than with 90 degrees and 180 degrees SLT treatments. 90 degrees SLT is generally not effective. 360 degrees SLT appears to be an effective treatment with approximately 60% of eyes achieving an IOP reduction of 30% or more. Transient anterior uveitis with associated ocular discomfort is not unusual in the first few days after SLT. Late complications causing ocular morbidity after SLT were not encountered.

Xeroderma pigmentosum (case report).

Xeroderma pigmentosum is a rare, hereditary and fatal disease of the skin. Ocular involvement is known to occur in 80% of cases. A case with typical cutaneous and ocular lesions is reported.