Leukocytes Of Patients Research Articles

Chronic granulomatous disease in an adult male: A proposed X-linked defect

A 25-year old patient with chronic granulomatous disease of somewhat unusual history is described. The diagnosis of CGD was based on increased susceptibility to infection, granulomatous appearance of tissues, and diminished bactericidal and metabolic response of leukocytes during phagocytosis: the clinical and cellular features considered phenotypic of CGD. A 16-year-old female sibling had bactericidal and metabolic abnormalities of leukocyte function similar to those of the patient's leukocytes. Leukocytes from another sister, 26 years of age, were intermediate in bactericidal capacity. Two populations of leukocytes were identified by a histochemical test of NBT reduction. Both normal and abnormal polymorphonuclear leukocytes were present in the leukocyte population of the two sisters. Leukocytes from the patient's mother and maternal grandmother were normal by all methods tested. These findings are taken as evidence of a germ-line mutation in the chromosomal gene causing CGD, with transmission of the genetic defect from the mother to the son.

Renal arteriography following systemic reaction to contrast material

A 15-year-old atopic female with severe hypertension (B.P. 220/160) was evaluated for evidence of renal arterial disease with equivocal results, and the performance of renal arteriography was considered essential. However, she had a history of two well-documented anaphylaxis-like reactions to contrast materials, considered to be an absolute contraindication to the performance of such a study. When the etiology of this sensitivity was investigated, an IgE-dependent mechanism was not evident. However, the patient's leukocytes released significantly greater quantities of histamine upon incubation with contrast materials than did 7 of 9 normal volunteers. The 2 hyperresponsive normal subjects were not atopic and had no prior exposure to contrast materials. Pretreatment with 80 mg prednisone and 200 mg diphenhydramine daily for 3 days prior to arteriography and challenge with increasing quantities of intravenous iothalamate meglumine (Conray) at the time of the study resulted in successful performance of arteriography and identification of fibromuscular dysplasia of the right renal artery. Placement of an aortorenal bypass graft corrected her hypertension. Thus, arteriography can be safely performed in selected patients with severe previous reactions to contrast materials when the alternatives provide no lesser risk by pretreatment with a steroid-antihistamine regimen and gradual administration of increasing doses of the contrast agent.

New Kind of Cytoplasmic Inclusions of Plasma Cells in Acid Maltase Deficiency

Extensive 'vacuolization' could be demonstrated in nearly all plasma cells and in some lymphocytes of an adult with glycogenosis type II (Pompe's disease). The biochemically defined diagnosis acid maltase deficiency (AMD) could be ascertained by examination of the maltase activity of the patient's leukocytes. Electron microscopical, microspectrographic, and cytochemical investigations revealed electron dense inclusions, which show an UV absorption at 276 nm and a positive reaction after PAS staining of plastic embedded material. Since no other abnormalities of the plasma cells could be found, our results are presumably indicative for a connection of AMD and a glycoprotein storage in the plasma cells of the patient.

Infusion of normal HL-A identical leukocytes in Sanfilippo disease type B. Estimate of infused cell survival by assays of alpha-N-acetylglucosaminidase activity and cytogenetic techniques: effect on glycosaminoglycan excretion in the urine.

A 101/2-year-old patient with Sanfilippo syndrome type B, after moderate immunosuppression, was infused with 2.2×10 10 leukocytes from his normal histocompatible brother. The patient's leukocytes lacked α- N -acetylglucosaminidase activity, while in the donor's cells this activity was in the range for homozygous normal individuals. Cultured patient and donor lymphocytes could also be distinguished on the basis of different chromosome fluorescence staining patterns. Both enzymatic assays and cytogenetic studies indicated that, for the first three weeks after infusion, 20% to 40% of circulating leukocytes had been derived from the donor. After the infusion there was a transient increase in the ratio of small to large glycosaminoglycans in the urine, without change in the clinical status. There was no long-term improvement in the clinical or biochemical manifestations of the disease.

Monocyte production of colony stimulating factor in familial cyclic neutropenia.

Summary. Granulocyte‐macrophage colony formation in agar culture of marrow is dependent on the presence of a glycoprotein colony stimulating factor (CSF). Measurement of the plasma levels of this factor in a 15‐yr‐old child with familial cyclic neutropenia revealed regular cycles with peak levels coinciding with nadirs of neutrophils and peaks of monocytes. A close correlation was observed between the incidence of monocytes and the capacity of the patient's leucocytes to elaborate CSF in culture. The frequency and amplitude of the blood monocyte cycles together with in vitro measurements of monocyte CSF‐producing capacity adequately accounted for the observed fluctuations of plasma CSF levels. A model of granulopoiesis is presented in which a positive stimulus for granulopoiesis is provided by CSF elaborated by nionocytes and a negative or neutral influence exerted by mature granulocytes.

Leukocyte Glucocerebrosidase Deficiency Diagnostic in Adult Gaucher's Disease with Negative Bone Marrow Biopsy. Some Properties of the Enzyme in Leukocytes and Spleen

Abstract. A sixty‐one year‐old man with Gaucher's disease is described. The disease was manifested by splenomegaly, hypersplenism and elevations of serum acid phosphatase and immunoglobulin but no bone lesions were roentgenologically demonstrable and bone marrow biopsy failed to reveal Gaucher cells. The diagnosis was established prior to splenectomy by the finding of decreased β‐D‐glucoside glucohydrolase (β‐D‐GGH) activity in his peripheral leukocytes. Examination of the surgically removed spleen revealed Gaucher cells, a large excess of glucocerebroside and decreased β‐D‐GGH activity. Splenectomy was followed by normalization of haemoglobin, white cell and platelet counts and of the serum acid phosphatase and immunoglobulin levels. The patient's leukocyte β‐D‐GGH activity showed an optimum pH similar to that of normal leukocyte enzyme, about 5.4, using both glucocerebroside (Glc‐Cer) and 4‐methylumbelliferyl‐β‐D‐glucopyranoside (MU‐Glc) as substrate. The patient's spleen β‐D‐GGH activity showed a pH optimum of about 4.5, which differed slightly from that of normal spleen enzyme. The Km values of β‐D‐GGH in the patient's leukocytes for Glc‐Cer and MU‐Glc were 1.35 × 10−4 M and 3.6 × 10−3 M, respectively, i.e. in the normal range. The patient's splenic tissue Km values were for Glc‐Cer and MU‐Glc 0.52 × 10−4 M and 5.8 × 10−3 M, respectively, as compared to 0.30 × 10−4 M and 5.0 × 10−3 M in a normal control.

PYROGLUTAMIC ACIDURIA Studies in an Infant with Chronic Metabolic Acidosis

SummaryA one‐year‐old girl with chronic metabolic acidosis was found to have normal renal acidi‐fication. In the urine, however, she excreted a daily amount of 50 mmoles of L‐pyroglutamate (5‐oxo‐˜‐proline). This excretion did not increase with increasing protein intake. Studies of the turnover of L‐pyroglutamate under steady state conditions revealed that 75% of the pyroglutamate synthesized was metabolized by the patient. The level of pyroglutamate degrading enzyme (5‐0times0‐prolinase) in the patient's leucocytes was not decreased, and the enzyme appeared to have normal affinity for its substrates, i.e. pyroglutamate and ATP.

IMMUNODIAGNOSTIC TEST FOR TRANSPLANT REJECTION

A simple, rapid, and specific immuno-logical test has detected the onset and progression of the immune responses of patients to their renal allografts. Within 5 hours of their application, immune (but not non-immune) peripheral leucocytes aggregated around donor-type kidney or skin fibroblast cells which had been maintained as monolayers in tissue culture. The pattern of reactivity in twenty-three patients revealed immunological specificity for antigens present in the kidney donor. Leucocyte aggregation in vitro was detected several days before clinical evidence of rejection. After reversal of rejection, host peripheral leucocytes were unresponsive in vitro. On the other hand, if the rejection could not be reversed and the transplanted kidney was removed, the patient's leucocytes displayed specific cellular reactivity for months. These observations suggest that an in-vitro test may be used in the routine evaluation of graft recipients to detect pre-existent sensitisation and to obtain an early diagnosis of renal allograft rejection.

TREATMENT OF MUCOCUTANEOUS CANDIDIASIS WITH TRANSFER FACTOR

Evaluation of a patient with chronic mucocutaneous candidiasis revealed that the patient lacked delayed hypersensitivity to candida and other skin test antigens and the patient's lymphocytes could not be stimulated in vitro with candida antigen. Phytohemagglutinin stimulation of lymphocytes in vitro was normal. Candidacidal function and myeloperoxidase activity of the patient's leukocytes were normal. The patient's lymphocytes produced migration inhibitory factor in response to candida antigen and the monocyte receptor sites for IgG were intact. Repeated treatments with amphotericin B, alone or with 5-fluorocytosine, were followed by immediate relapse. Four doses of transfer factor were administered and when coupled with amphotericin B on two occasions prompted remissions of 6 and 4 months, respectively.

Biologic properties of E myeloma proteins

E myeloma protein PS (PS protein) from the serum of the second known patient (P.S.) with IgE myeloma was purified by DEAE-cellulose column chromatography and Sephadex ® G-200 gel filtration. The physicochemical properties of this protein were identical with those of E myeloma protein ND (ND protein) from the first patient with IgE myeloma; however, double immunodiffusion studies revealed distinct idiotypic antigenic determinants probably belonging to the Fd portion of the molecule in both E myeloma proteins. Patient P.S. failed to accept passive sensitization with reaginic antibodies (the Prausnitz-Kustner reaction) and reacted only to the intradermal injections of high concentrations of antilgE (the reversed Prausnitz-Kustner reaction). He was capable of reacting normally to the intradermal injection of histamine. It is postulated that the absence of passive sensitization with reaginic antibodies was due to saturation of IgE-fixing sites of target cells with E myeloma protein. The impaired reversed Prausnitz-Kustner reaction may be explained by failure of the injected anti-IgE to react with cell-fixed IgE because of the presence of an excessive amount of E myeloma protein in extracellular fluid. Histamine release from the patient's leukocytes by treatment with anti-IgE was studied. Studies of IgE metabolism were also made, using 125I-PS protein and 131I-ND protein. The metabolic parameters of both proteins were identical, with a half-time of 5.1 days and fractional catabolic rate of 16 per cent in patient P.S., in contrast to 2.5 days and 89 per cent, respectively, in normal subjects.

VITAMIN B12-DEPENDENT METHYLMALONICACIDURIA: AMINO ACID TOXICITY, LONG CHAIN KETONURIA, AND PROTECTIVE EFFECT OF VITAMIN B12

Methylmalonicaciduria is an inborn error of metabolism characterized by protein intolerance, neonatal or infantile ketoacidosis, long chain ketonuria, and by the accumulation of large amounts of methylmalonic acid (MMA) in blood and urine. The present study examines the metabolic basis for these features in a young boy with the vitamin B12-dependent variant of this disorder. A diet containing 5 gm of protein per kg body weight per day resulted in transient ketoacidosis, long chain ketonuria, and increased methylmalonicaciduria. Isoleucine, valine, methionine, and threonine, all known MMA precursors, were responsible for this protein intolerance. Oral loads of each of these amino acids resulted in equimolar 1,000 µg increases in urinary MMA. Valine and isoleucine also precipitated mild ketoacidosis and long chain ketonuria. Injections of vitamin B12 (1000 µg per day) promptly lowered urine MMA by 70% and partially corrected the defect in propionate or methylmalonate oxidation found in the patient's leucocytes or platelets. While receiving vitamin B12, the clinical toxicity of a valine load was abolished and its biochemical effects significantly modified. These findings demonstrate that the observed protein intolerance and ketoacidosis in patients with methylmalonicaciduria are direct results of the primary block in methylmalonate catabolism and that vitamin B12 acts by overcoming, in part, this metabolic block. These conclusions imply that children with this disorder should respond to dietary protein restriction and/or vitamin B12 supplementation, a regimen on which our patient has thrived for over 2 years.

Chronic Myelogenous Leukemia: Cyclic Leukocytosis and Identical Twin Discordance

A cyclic leukocytosis with cycles of approximately 72 days and counts ranging from 4,000 to 145,000 cells/mm3 has been followed for 15 months with no treatment, in a 13-year-old white female with chronic myelogenous leukemia (Ph1 positive). An identical twin has no clinical, morphologic or cytogenetic evidence of leukemia. Differential counts of per. bl. and marrow remain unchanged during the patient's cycles although marrow cellularity is increased during peaks. Studies which were normal include: Rebuck skin windows, phagocytic and bactericidal functions of neutrophils, glutathione peroxidase levels, fetal hemoglobin and immunologic responses. Epinephrine stimulation produced a total release of marginal pool ceels at the high point with only partial release of cells at a low point. Endotoxin and steroid stimulation studies indicate that mechanisms for delivery of cells from marrow are intact and of similar magnitude at both high and low points. Ph1 positive cells were seen in 81% of mitoses from per. bl. cells taken at a peak while in per. bl. specimens taken at a low point such Ph1 positive cells were absent. At both high and low points over 90% of mitoses of bone marrow cells were Ph1 positive. mixed leukocyte cultures showed a non-reciprocal response of the patient's leukocytes to cells of her identical twin, suggesting partial loss of transplantation antigens in some of the patient's cells. Leukokinetics with DFP32-labeled cells, alkaline phosphatase and granulopoietin levels and marrow proliferation studies with H3T were measured at high and low points. These studies would be compatible with the view that controls of leukocyte production and release as well as in immunogenetic characteristics of cells are involved in this leukemic process.

Studies on Alkaline Phosphatase in Hypophosphatasia

Central to an understanding of the pathogenesis of hypophosphatasia is the question of whether or not there is a deficiency of alkaline phosphatase (AP) activity in bone. A recently reported patient with the full syndrome had consistently normal total AP activity in serum and the suggestion was made that deficiency of phosphatase was not of primary pathogenic importance [SCRIVER and CAMERON, NEW Engl.J.Med. 281: 684, 1969].We have studied two children who have the typical osseous and dental lesions of hypophosphatasia, markedly increased excretion of phosphoethanolamine, and total activity of serum AP in the normal adult range. AP was undetectable histochemically in leukocytes of the patients and was normal in family members. Extraction of the patient's leukocytes yielded extremely low activity. Electrophoresis of the patients' sera on polyacrylamide gel consistently yielded a single major band of phophatase activity with a rate of migration intermediate between the rates of the two fast bands seen in normal sera. The two fast bands in normal sera have been identified as hepatic and osseous AP by comparison with extracts of tissues. Although the dominant band in the patients appeared to migrate differently from any seen in normal sera, there were no unusual isozyme patterns of AP from sera and leukoscytes of family memebers. Studies of sensitivities of various AP to heat, 1-phenylalanine, and urea did not clearly distinguish the patient's sera from normal sera, nor did these studies indicate clearly the tissues sources of serum enzymes. We conclude that abnormalities of AP do exist in hypophosphatasia, and that most of the AP in sera of patients is derived from tissues other than bone.

Leucocyte alloantigens in the kidney. I. Serological identification.

Summary Glomeruli have been prepared from 12 sets of human cadaver kidneys. They have been used to absorb polyspecific leucocyte alloantisera, and leucocyte alloantigens have been demonstrated in 11 out of 12 sets of glomeruli. In three cases, alloantigens detected on the patient's leucocytes were shown to be present in the glomeruli. In one case, the glomeruli contained, in addition, an antigen not detected on the leucocytes by cytotoxicity. In one case, using a monospecific alloantiserum, the antigen T12 was detected on leucocytes and glomeruli.

Facial Granuloma Associated With Fusarium Infection

A teen-aged boy developed a facial granuloma due to the saprophyte<i>Fusarium</i>. Since no similar case has been reported to our knowledge, it suggests that abnormal features are involved. Th was borne out when it was discovered that the patient's leukocytes functioned imperfectly. He is a member of a group of patients with "chronic granulomatous disease."

Hyperglycinemia and propionate carboxylation.

The oxidation of propionate and methylmalonate was investigated in leukocytes from a patient with hyperglycinemia. It was found that the patient's leukocytes could oxidize both propionate and methylmalonate normally. These findings suggest that hyperglycinemia consists of primary type and secondary type.

Increased lecithin synthesis during phagocytosis by normal leukocytes and by leukocytes of a patient with chronic granulomatous disease.

Study of incorporation of albumin-bound lysolecithin-32P into lecithin of human leukocytes (from peripheral blood) during ingestion of inert particles revealed that significantly more lysolecithin was incorporated than at rest. This stimulation of lecithin synthesis accounts for substantial addition of new lecithin and may be important for membrane formation during phagocytosis. Leukocytes from a patient with chronic granulomatous disease, cells that are known to manifest decreased bactericidal activity but normal phagocytosis, also exhibited increased lecithin synthesis, during engulfment of particles in vitro, from lysolecithin in the medium. Electron microscopy of the patient's leukocytes showed formation of phagosomes and degranulation indistinguishable from similar phenomena in leukocytes from normal persons.

Iodination defect in the leukocytes of a patient with chronic granulomatous disease of childhood.

In a case of chronic granulomatous disease, the patient's neutrophils, unlike normal neutrophils, did not fix iodine after ingestion of heat-killed Lactobacillus acidophilus or heat-killed or live Serratia marcescens. When live L. acidophilus was employed, iodination did occur, although it was less than normal. The patient's mother had some neutrophils that did not and some neutrophils that did iodinate after ingestion of heat-killed L. acidophilus. Unlike S. marcescens, L. acidophilus was killed as well by the patient's leukocytes as by normal leukocytes. Methimazole inhibited the killing of L. acidophilus by the leukocytes of the patient and the patient's mother but had no significant effect when normal leukocytes were employed. The ability of leukocytes in chronic granulomatous disease to kill certain organisms may depend on bacterial formation of the hydrogen peroxide required for an antimicrobial system in the leukocytes, mediated by myeloperoxidase.

31 The 'Impotent Neutrophil' Syndrome

Leukocyte function was studied in a 2–2,5-year-old Negro male with the history of repeated abscesses, cervical and inguinal adenitis, recurrent pneumonia, seborrheic eczema of the scalp, hepatosplenomegaly and anemia. An appropriate leukocytosis with neutrophilia occurred with each infection, absolute neutrophil counts ranging from 3,900 to 24,500/mm3. IgG, IgA and IgM globulins were present in increased amounts. Lymph node biopsy contained caseating granuloma. Skin tests and cultures for all types of acid fast bacilli and fungi were repeatedly negative. These findings are characteristic of the syndrome of chronic granulomatous disease of childhood. There was a normal flux of leukocytes into Rebuck skin windows and exudate fluid. Leukocytes appeared normal with alkaline phosphatase, peroxidase and Wright's stain and were normally vacuolated after bacterial phagocytosis. Normal leukocyte motility and phagocytosis followed by degranulation were seen with phase microscopy. Decreased bactericidal activity of the patient's leukocytes during in vitro incubation with Staphylococcus aureus and Aerobacter aerogenes was repeatedly demonstrated in the presence of the patient's serum or normal serum. To determine the nature of this defect a citric acid extract of the patient's leukocyte granules was compared with control specimens and decreased bactericidal activity was found. The deficient or abnormal phagocytin activity of the patient's leukocytes, demonstrated by this study, could account for his clinical syndrome. (SPR)

CHROMOSOME ABERRATIONS IN LEUCOCYTES OF PATIENTS WITH ASEPTIC MENINGITIS.

Three patients with aseptic meningitis and two patients with scarlet fever were subjected to chromosome analysis in cultured leucocytes. Information was obtained that leucocytes of the aseptic meningitis patients exhibit random and non-specific chromosome aberrations significantly higher in frequency than the control series from scarlet fever patients. The most common type of the aberration observed was a single chromatid break including a full break in one chromatid. Isochromatid-type breaks, chromatid- and chromosome-type interchanges and acentric fragments were observed less frequently.