Shmuel Shtrasburg

The Hypereosinophilic Syndrome Associated with CD4+CD3″ Helper Type 2 (Th2) Lymphocytes

We describe herein the clinical and laboratory manifestations of a unique group of patients (pts) presenting with hypereosinophilic syndrome (HES) who were treated in our medical centers for 4–13 years. Skin biopsies, flow cytometry of peripheral blood mononuclear cells (PBMC), assays for cytokines and immunoglobulin (Ig) production in vitro, and Southern blots of T-cell receptor (TCR) genes were performed. All four pts had a persistent hypereosinophilia (>1.9 × 109/L) and chronic skin rash. Three of four had elevated IgE, thrombotic manifestations and lung involvement (asthma and/or infiltrates), and one had deforming sero-negative arthritis of the hands. 66–95% of their peripheral T-cells expressed CD4 but not CD3 or TCR molecules on the cell surface membrane. Activated CD4+CD3- cells secreted interleukin (IL)- 4 and/or 5, and were required for maximal IgE secretion by autologous B-cells. Two pts had evidence of rearrangement of TCR genes of the CD4+CD3- cells, one of whom died of anaplastic lymphoma. In conclusion, HES with CD4+CD3- lymphocytosis may be associated with high serum IgE, dermatological, pulmonary, thrombotic and rheumatic manifestations which may be due to Th2 effects of CD4+CD3-cells migrating to end organs. Fatal systemic lymphoid malignancy may also develop in some pts with monoclonal expansion of the CD4+CD3- T-cells.

Characterization of High Molecular Weight Amyloid A Proteins

Human amyloid A protein (AA) is usually composed of the NH2‐terminal 76 amino acid residue of serum amyloid A protein (SAA), although lower and higher molecular weight fragments have been reported. We studied the primary structure of six AA proteins with molecular weights of 11 kDA‐15kDA. as determined by SDS‐PAGE. Automated Edman degradation of the intact purified proteins and sequence analysis of enzymatic peptides revealed that the AA proteins were composed of only 74 to 87 residues. Moreover, fragments of apolipoprotein E or histones 2a, 3 and 4 were associated with these AA molecules. Thus, AA heterogeneity may reflect diverse processing of the SAA precursor and a very close association with other proteins.

Direct Evidence for SAA Deposition in Tissues During Murine Amyloidogenesis

To study the mechanism of amyloid deposition, the nature of amyloid proteins formed in experimental murine amyloidosis, was examined. Spleen specimens, 15–60mg, were homogenized and extracted using aqueous acidic acetonitrile, in a recently developed procedure, making it possible to obtain amyloid proteins from minute amounts of tissue. The extracted material, 1.5–4mg, was analysed by Western blotting and ELISA using antibodies recognizing differentially proteins AA and SAA. Two immunoreactive proteins of 8 and 12 KDa were isolated and subjected to amino acid analysis and N‐terminal sequence determination. The results of immunochemical and chemical examination showed that the 8 and 12 KDa proteins represented proteins AA and SAA, respectively.

Crush and Smear Technique for Rapid Detection and Semiquantitation of Amyloid Deposition

A method using Congo red to rapidly identify and semiquantitate amyloid deposits in tissues for experimental research and clinical medicine is described. Examination by polarization microscopy revealed amyloid deposits as bright green birefringent clumps on a dark red background. On semiquantitative evaluation, good correlation was found between this technique and the conventional histological one, the present technique being more sensitive. The method described saves time and expense.

Pregnancy and amyloidogenesis : I. Offspring of amyloidotic mice are not predisposed to develop amyloidosis

Amyloid enhancing factor (AEF) is a substance formed during amyloidogenesis that accelerates amyloid deposition in tissues. The administration of AEF followed by AgNO3 (inflammatory stimulus) to mice results in amyloidosis within 6 days. The purpose of the study was to determine whether the offspring of amyloidotic mice are exposed to maternal AEF during pregnancy and therefore become predisposed to the development of amyloidosis on inflammatory stimulus. To that end female mice were made amyloidotic by the administration of AEF and AgNO3, made pre-amyloidotic (a condition associated with self-generation of AEF) with a short course of casein, or treated with exogenous AEF without AgNO3; then mating and conception took place. The possible priming of offspring with maternal AEF was studied by the administration of AgNO3 alone (without AEF) to the offspring and the determination of the presence of amyloid deposits in their spleens. Despite the long-term effect of AEF and its high activity, amyloidosis did not develop in any of the studied offspring, implying that the newborn mice were not primed by maternal AEF. These findings suggest that amyloidotic mothers do not predispose their offspring to the risk of developing amyloidosis, probably because maternal AEF does not cross the placenta.

Amyloid goiter: report of the clinical, histological and biochemical features of five cases

The clinical course and pathological findings of five cases of amyloid goiter in patients with AA amyloidosis are presented. The underlying diseases which caused the amyloidosis were: chronic recurrent multifocal sterile osteomyelitis, acne conglobata, tuberculosis, and two cases of familial Mediterranean fever. All presented with a hard, moderately rapidly growing thyroid mass, mimicking a malignant thyroid tumor. Four patients underwent excisiorial biopsy or partial thyroidectomy, after which they developed hypothyroidism. The fifth case was discovered at an autopsy.Histologically, in all cases, the thyroid follicles were scarce, many had an atrophic appearance, surrounded by amyloid deposits and associated with adipose metaplasia of the stroma. The amyloid was potassium permanganate Congo red sensitive, and reacted with anti-AA antibodies by the avidin biotin immunoperoxidase method in all cases.The amyloid fibrils, extracted from the thyroid tissue of each of the five cases, revealed two protein bands...

Systemic amyloidosis in a cheetah (Acinonyx jubatus)

A captive, 9 year old female cheetah (Acinonyxjubatus) with progressive signs of chronic renal failure was euthanized Histologic studies of the kidney revealed severe glomerular and medullary amyloidosis. The amyloid stained positivelv with Congo red and showed green birefringence under polarized light. Based on potassium permanganate sensitivity and the immunohistochemical reaction with monoclonal antibodies, the renal amyloid was classlfied as AA. Amyloid was also deposited in the liver and heart. Elec-trophoresis of heart and kidney tissue showed two protein bands reflecting serum amyloid A. This is the first detailed report of amyloidosis leading to renal failure in a highly endangered species.

Quantification of tissue amyloid content in AA amyloidosis by inhibition ELISA

The determination of the amount of amyloid in tissues is important to follow the onset and the development of amyloidosis. In the present study a new procedure for quantification of amyloid proteins in AA amyloidosis is described. It involves the protein extraction from mouse and human tissue and evaluation of their amount by inhibition ELISA. The procedure is simple and may be performed using milligram amounts of tissue. The sensitivity limit of protein AA detection corresponds to 0.007mg/mg tissue. The results obtained using the inhibition ELISA were consistent with those obtained by Congo red staining. In contrast to previously developed amyloid quantification techniques, the presented procedure required only the basic equipment avail-able in most laboratories.

Extremely high dose pravastatin may suppress amyloidogenesis in a mouse model

Pravastatin is a cholesterol lowering agent,1 recently reported to have anti-inflammatory properties.2 It was suggested that the prevention and regression of atherosclerosis by pravastatin is partially related to its anti-inflammatory effect,2 probably mediated by inhibition of proinflammatory cytokines.3 Pravastatins anti-inflammatory effect is associated with, and probably reflected by, reduced levels of the acute phase reactants, C reactive protein and serum amyloid A (SAA).2 The N-terminal fragment of the SAA is amyloid A (AA), which is deposited in a fibrillar form in the tissues of up to 30% of patients with a variety of …