Profiling the immune responses of human patients treated with recombinant streptokinase after myocardial infarct.
The SPOT synthesis of peptide arrays on continuous cellulose membranes should be generally applicable in the analysis of sequential antibody binding sites using the enzyme-substrate or other standard detection protocols. The use of total serum is limited by the occurrence of high background levels. This may be overcome if affinity purified antibodies or sera with high antibody titers are used, which allows work at high dilutions and a consequent reduction of background level. Here we demonstrate the mapping of antigenic regions located on recombinant streptokinase SK-2 (Heberkinase) using cellulose-bound peptide scans and human total sera from patients treated with SK-2 (Heberkinase). Streptokinase (SK) is a 47 kDa protein produced by various strains of hemolytic streptococci and is a potent activator of the fibrinolytic enzyme system in humans. SK is in widespread clinical use to treat acute infarction because of its function as an activator of vascular fibrinolysis. Since streptococcal infections are common, normal individuals are immunized with SK and antibodies (Abs) to SK can be detected in most of them. This therapy generates significant T-cell responses to SK and the neutralizing capacity of the Abs rises significantly. Neutralizing Abs reduces the efficiency of thrombolytic therapy and may cause allergic reactions. The widespread use of SK in humans makes its antigenicity an important clinical problem. In this regard the study of the immunodominant regions of SK becomes an important aspect for the improvement of this thrombolytic agent.
Reyes
O
O
Centro de Ingeniería Genética y Biotecnología, P.O. Box 6162, Havana, Cuba. oreyes@cigb.edu.cu
Torrens
I
I
Ojalvo
A G
AG
Seralena
A
A
Garay
H E
HE
D000595:Amino Acid Sequence; D005343:Fibrinolytic Agents / Q000627:therapeutic use*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009203:Myocardial Infarction / Q000188:drug therapy* / Q000276:immunology*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry; D019151:Peptide Library; D040081:Protein Array Analysis; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000627:therapeutic use; D013300:Streptokinase / Q000737:chemistry / Q000627:therapeutic use*
D005343:Fibrinolytic Agents; D010446:Peptide Fragments; D019151:Peptide Library; D011994:Recombinant Proteins; D013300:Streptokinase