Identification of candidate target antigens for antibody-based immunotherapy in childhood B-cell precursor ALL.

BACKGROUND: Contemporary risk adapted treatment protocols for childhood acute lymphoblastic leukemia (ALL) rely on accurate risk assessment strategies for disease re-occurrence by incorporating clinical parameters as well as immunological, molecular and cytogenetic features of the blasts at initial manifestation. Additional risk stratification is provided by analysis of the IN VITRO and IN VIVO response of the blasts towards standard chemotherapy. Despite adapted therapies, a number of children with good and bad prognostic factors still fail therapy. One approach to this problem might be to incorporate monoclonal antibodies (MoAbs) as additional modalities into the first or second line treatment. PATIENTS AND METHODS: In order to identify target antigen structures, we analyzed the immunological expression profiles of blasts from 181 patients with B-cell precursor ALL treated at our institution in 11 years according to the CoALL-92/97/03 protocols. Blasts were classified according to the EGIL guidelines as 9 proB-, 110 common (c-) and 62 preB-ALL. RESULTS: > 99 and 96 % of patients expressed CD19 and CD22 on > 90 % of their blasts, respectively. HLA-DR on > 95 % blasts was present in all patients. CD10 was expressed on all c-/preB-ALL and absent on proB-ALL cells. CD20 was expressed on 11-37 % of B-cell precursor ALL samples. CD34 positive blasts were found in 89, 83 and 68 % of patients with proB-, c- and preB-ALL, respectively. CD37 expression was detected in 0-18 % of patients. < 20 % CD45(+) blasts were found in 11, 19 and 18 % of patients with proB-, c- and preB-ALL. CD33(+) was expressed on 33, 29 and 21 % of patients samples with proB-, c- and preB-ALL. Other myeloid antigens (CD13, CD14, CD15, CD65) were positive on blasts in < 25 % of patients. Analyses of the immunological profile of blasts in 9 consecutive children with relapse revealed that the antigen expression profile varied little compared to the initial diagnosis for CD10, CD19, CD22 and HLA-DR. CONCLUSIONS: These analyses clearly identified the three antigens CD19, CD22 and HLA-DR present on blasts in more than 90 % of patients as potential target structures for targeted therapies with native or toxin-bound monoclonal antibodies in childhood ALL.

[In vitro susceptibility to antifungal agents of Candida strains isolated from patients with various diseases of the respiratory tract]. / Wrazliwosc na leki przeciwgrzybicze in vitro szczepów Candida wyizolowanych od chorych na rózne choroby ukladu oddechowego.

The aim of the study was the estimation of in vitro susceptibility to antifungal agents of yeast isolated from sputum of 70 respiratory diseases patients using the disc-diffusion method-antimycogram. The following agents were tested: amphotericin B, 5-fluorocytosine, nystatin, ketoconazole, fluconazole. Only Candida strains were isolated from sputum, 82% of them were Candida albicans. We noted differences in susceptibility to antimycotics of Candida strains. The best antimycotic in vitro was 5-fluorocytosine. 54% of isolated Candida strains were resistant to 1 or more antimycotics.

[Evaluation of occurrence of fungal infection in patients with lung cancer]. / Ocena wystepowania zakazenia grzybiczego u chorych z rakiem pluc.

The study was aimed at determination of frequency and size of fungal infection in patients with lung carcinoma, as compared with the control group. The study involved 20 patients with histologically confirmed lung carcinoma and--as a control group--20 patients with other lung diseases and healthy volunteers. Quantitative mycological investigation of sputum and bronchial rinsings were performed threefold. The degree of infection of these materials was determined on a Sabouraud medium. Candida-like fungi were determined basing on API 20 AUX test. In majority of patients with lung carcinoma, presence of pathogenic fungi, mostly from the genus Candida, was detected in sputum or bronchial secretions. In one third of cases infection was severe or very severe which was dependent of the stage of disease advancement. In control group severe infection was observed only sporadically.

[Trial of evaluating co-existent fungal infection during course of neoplastic disease in patients with lung cancer]. / Próba oceny wplywu wspólistniejacego zakazenia grzybiczego na przebieg choroby nowotworowej u chorych z rakiem pluc.

The study was aimed at determination of possible connection between co-existing fungal infection in patients with lung carcinoma with course of neoplastic disease and ability to achieve remission by application of different methods of treatment. The study involved 25 patients with histologically confirmed lung carcinoma. Their sputum and bronchial rinsings were tested mycologically. The tests were performed at the moment of diagnosis of the diseases, during and after treatment. In all patients occurrence and type of Candida infections was investigated. Dynamics of fungal infection was measured by a scale from minimal to very severe. In about 1/3 of patients at the moment of diagnosis of the disease, severe or very severe infection was seen which was dependent of stage of the disease advancement. These preliminary studies seem to indicate that presence of severe or very severe fungal infection is not favourable prognostically and frequently correlates with a progress of neoplastic changes and worse response to treatment.

[Influence of treating hypothyroidism and hyperthyroidism upon psychical reaction time]. / Wplyw leczenia niedoczynnosci i nadczynnosci tarczycy na czas reakcji psychicznej.

The aim of the study was answering the question whether determination of psychical reaction time may be useful for the monitoring of treatment of hypothyreosis and hyperthyreosis. The correlation of disease symptoms (diagnostic index for hypothyroidism after Murray and for hypothyroidism according to Crooks), concentration of triiodothyronine, thyroxine, index of free thyroxine in blood serum and Achilles tendon reflex--with the simple reaction time (srt) as well as with the choice reaction time (chrt) before and during treatment was investigated in 18 patients with primary hypothyroidism and in 24 with hyperthyroidism caused by Graves-Basedow's disease. Two control groups comprised 84 healthy persons. Either in hypothyroidism or in hyperthyroidism the srt and chrt was significantly prolonged. The substitution therapy in hypothyroidism resulted in a normalization of both parameters. On the other hand, administration of thiamazole, especially in the early period of treatment of hyperthyroidism, further prolonged the srt and chrt. A shortening of the time of each of the two reactions occurred, however, when the restoration of euthyroidism was achieved, which was accompanied by reduction of thiamazole dosage. The obtained data point to the usefulness of srt and chrt determination for the diagnosis and therapy monitoring of functional disturbances of the thyroid gland.

[Treatment of sudden deafness with xantinolnicotinate (Complamin) in increasing dosage]. / Zur Behandlung des Hörsturzes mit Xantinolnicotinat (Complamin) in aufsteigender Dosierung.

Twenty-nine patients affected by typical unilateral sudden hearing loss were treated with Xantinolnicotinate (Complamin) by a 21 days infusion of 1.5 g-30 g in 500 ml NaCl 0.9% followed by 3 months 1,000 mg sustained release tablets t.i.d. An improvement in hearing was achieved in 90.5% of the cases and in 52% returned to normal. Tinnitus improved in 92% of cases and resolved completely in 54%. The results of speech and sound audiograms correlated well, and showed significant recovery in the first 3 weeks. The infusion therapy was terminated prematurely in 4 patients because of near normalization of hearing (maximum dosage: 15 g Complamin). Adverse reactions only occurred during the infusion therapy. For ethical and therapeutic reasons a placebo control group was not included. To compensate for this the audiograms of impaired and 'normal' ears were compared. The changes in hearing and tinnitus are therefore to be regarded as the result of spontaneous remission and drug efficacy.

Determination of femtomol quantities of gibberellic acid by radioimmunoassay.

A highly sensitive and specific radioimmunoassay which allows the detection of as little as 5 fmol (2 pg) of gibberellic acid (GA3) in crude plant extracts is described. Antisera of high affinity and titer were obtained by immunizing rabbits with a conjugate of carboxyl-coupled GA3 and bovine serum albumin. [(125)I]Gibberellic acid-[N-(p-hydroxybenzyl) putrescine]amide of high specific activity, used as the immunotracer, is readily displaced by gibberellic acid methyl ester but not by free gibberellic acid. Thus, methylation of extracts prior to analysis is required. The assay is very specific; besides GA3, only the closely related GA7 is highly immunoreactive. Various gibberellins, related compounds, as well as other classes of plant hormones do not interfere with the assay. Levels of immunoreactive gibberellins (GA3, GA7) in actively growing tissues, among them cell suspension cultures of 33 different species, were determined.