Cloning and expression of a Leishmania donovani gene instructed by a peptide isolated from major histocompatibility complex class II molecules of infected macrophages.

The studies reported here describe the isolation of peptides from MHC class II molecules of murine macrophages infected with Leishmania donovani, and the use of the derived peptide sequences to rescue the pathogen peptide donor protein. The isolation of the peptides was carried out by comparing the RP HPLC profile of peptides extracted from infected macrophages with the peptides extracted from noninfected cells. Several distinct HPLC peaks unique to infected macrophages were sequenced. One of the peptides that was not homologous to any known protein was used to instruct the designing of an oligonucleotide sense primer that was used in combination with an oligo dT nucleotide (anti-sense primer) to amplify by PCR a DNA fragment from L. donovani cDNA. The amplified DNA fragment was cloned and used as a probe to screen a L. donovani cDNA library. The cloned gene (Ld peptide gene) has an open reading frame of 525 bp and has no homology with any known protein/gene sequence. Northern blot analyses indicated that the Ld peptide/gene is broadly distributed and expressed among species of the Leishmania genus, in both the amastigote and promastigote life cycle forms. Using the pGEX 2T vector, the gene was expressed and the relationship of the purified recombinant protein with L. donovani was confirmed using both antibody and T cell responses from immunized or infected animals. The gene encodes a 23-kD molecule (Ldp 23) associated with the cell surface of L. donovani promastigotes. In addition, T cells purified from the lymph nodes of BALB/c mice immunized with L. donovani or infected with L. major, and from CBA/J mice infected with L. amazonensis were stimulated to proliferate by the recombinant Ldp 23 and produced high levels of IFN-gamma and no IL 4. This observation suggests that the Ldp 23 is an interesting parasite molecule for the studies concerning the host/parasite interaction because the Th1 pattern of cytokine response that it induces is correlated with resistance to Leishmania infections. These results clearly point to an alternative strategy for the purification of proteins useful for the development of both vaccines and immunological diagnostic tools not only against leishmaniasis but also for other diseases caused by intracellular pathogens.

[Multidisciplinary study of cerebrovascular disorders. II. Cardiovascular profile of occlusive vascular disorders]. / Estudio multidisciplinario de los accidentes vasculares encefálicos. II--Perfil cardiovascular de los accidentes vasculares oclusivos.

Ischemic stroke constitute a mayor cause of morbidity and mortality in the adult population, particularly in the elderly. Heart disease may predispose to ischemic stroke, especially in the presence of transient or permanent precipitating factors such as atrial fibrillation. To elucidate the role of heart disease in predisposing to ischemic stroke we studied the clinical and non invasive cardiac profile (EKG, 2D-Echo, Holter) of 186 consecutive patients, 91 of them embolic (GI) and 96 non embolic (lacunar, atherothrombotic, others) (GII), as determined by brain CT scan and thorough clinical evaluation. Age and male/female ratio were significantly different (71 + 13 vs 65 + 12 years, 40/60 vs 65/35, p < 0.003). Hypertension was equally common in both groups (38 and 40%). Patients in GI had higher prevalence of valvular heart disease (23 vs 1%), atrial fibrillation (67 vs 10%), 2D Echo left atrial enlargement (45 vs 16%) and supraventricular ectopy in Holter (59 vs 32%) p < 0,001. By contrast absence of heart disease (45 vs 19%), ST-T changes in EKG (28 vs 14%), left ventricular hypertrophy in 2D Echo (28 vs 9%) and ventricular ectopy in Holter (54 vs 23%) were more prevalent in GII patients, p < 0.001. Multiple stepwise logistic regression analysis showed age > 70 years (relative risk (RR) 1.67), valvular heart disease (RR 2.25), chronic AF (RR 2.44) and paroxysmal AF (RR 1.89) were significant independent predictors of embolic stroke, whereas the presence of left ventricular hypertrophy in 2D-Echo (RR 0.76) and frequent ventricular premature beats in Holter (RR 0.47) were predictors of occlusive non embolic stroke. Thus, the clinical and non invasive cardiac profile of embolic and non embolic ischemic stroke is significantly different, which is relevant to preventive strategies.

[Anticardiolipin antibodies and primary antiphospholipid syndrome of the central nervous system: report of 10 clinical cases]. / Anticuerpos anticardiolipinas y síndrome antifosfolípidos primario del sistema nervioso central: comunicación de 10 casos clínicos.

An association between anti-phospholipid antibodies and different disorders of the central nervous system has been described recently. We used an ELISA technique and detected anti-cardiolipin antibodies of the IgG or IgM variety in a series of 10 patients: 4 had occlusive stroke, 2 brain hemorrhage, 2 chorea, 1 a Sneddon syndrome and 1 vascular cephalea. The diagnosis of primary anti-phospholipid syndrome was suggested after ruling out systemic lupus in all patients.