T H E J OU RNA l. OF I NV ESTI GATI VE D E RMATOLOGY

Co p y righl © 1973 by The Willia ms & Wilkins Co. Vol. 60, No.6

Printed in U.S. A.

THE CHEDIAK- HIGASHI SYNDROME AND THE HOMOLOGOUS TRAIT IN ANIMALS*

DOROTHY B. WINDHORST, M.D.t AN D GEORGE PADGETT, D.V.M.

INTRO DUCTIO N

The Chedi a k- Higashi syndrome (CHS ) is in­cluded in a sympos ium on genetics and the skin be c a use of the unusual pigmentary diluti on that is a primary phenotypic ma nifestation of the condi ­tio n . Although the syndrome is rare in man , it ca n be investiga ted in severa l anima l homologues. The d isea se states, which are a part of t he syndrome, fasc ina te in vestigators of host defenses aga inst infections and malignancy, a nd the nat ure of the subcellula r morphologic defect offers considerable ch a llenge to t hose interested in cytopl as mic or­ga n e lles.

S e veral reviews of th e condi t ion (Windhorst et a l. , 1968a; Padget t , 1968; Padgett et a l. , 1970) have pro mpted this discussion which will focus on two a reas t hat wa rrant furth er comment : t he pigmen­tar y a nomaly and poss ible ex planations for t he suscept ibility to infect ious agents . First, a brief desc ript ion of the syndrome.

THE CLINI CA L STATE

Early Findings

C hildren brought in for treatment of CHS are gen e ra lly t hought by the phys ician to have some innocuous infect ion. Only if the pigmentary abnor­mali ty is recognized is t he t rue gravi ty of the syndrome rea lized . Higashi (1954) ment ioned t hat th e mother of his pat ient recogni zed t he dire pr ogn os is on t he bas is of her experience wi t h t hree pre vious s iblings who had had the sa me unusua l col o r. Thus, t he pigmenta ry anoma ly in a ll four children a lerte d Higashi to t he poss ibili ty that t he children were manifesting not just a se ries of m ul t iple infections but a fa mili a l t ra it. This sa me fa c tor may have cont ributed to t he prominence of early reports from count ri es wi t h populat ions con­sisting of rather uniformly dark-skinn ed individu ­als . Since t he CHS pigmentary defect is a regul a r dilut ion of other pigment factors rather than a total a bsence of co lor, it can be missed in a child of more heterogeneous pigmenta ry ba.ckground un­less t he physician t hinks in terms of t he individual pati e n t 's fa milial pigmentary constit ution (Wind ­hor st, 1968). In any event , a lt hough the earl y

T hi s work was supported in part by an America n Ca n cer Society Fac ul ty Scholar Award (DBW ) and Na t io na! Institutes of Hea lth Grants CA 05887, AI 06591 , and AI 06477.

* F rom the Department of' Medicine, Section of Der­mato logy, Pritzker School of Med icine, University of Chicago, Chi cago, lllinois, and the Departm ent of Veteri­nar y Pa thology, Washington State Uni versity , Pullman, Wash ington.

t P resent add ress: National Institutes of Hea lth, Building 10, Room 4B-l8, Bethesda, Maryland 20014.

529

descrip t ions by Beguez-Cesar (1943) and Stein­brinck (1948) included the pigmenta ry defect as a component of t he problem, Chedia k (1952) and particul arly Higas hi (1954) a rrived at t he synthesis of famili a l pigmen tary anomaly, suscept ibili ty to infect ion , morphologic a nomalies in the neutro­phils, and early death as an enti ty to which Higas hi 's mentor, Sato (1955), ult imately ap­pended. t he eponym.

Large Granules in Blood Smears

Well before the current understanding of lyso ­somes and t he theories of storage diseases , Higashi had ti t led his paper, almost prophetica lly, " Con­geni tal Gigant ism of Peroxidase Granules ." This t it le has stood the test of t ime; histochemica l studies of t he giant granules in blood and bone marrow prepara tions (Bessis et a l. , 1961 ; Mauri and Silinga rdi , 1964) showed them to be enla rged versions of t he usual granulat ion for a given cell ty pe ra ther than an accumulation of uniformly st aining materia l.

Poor Prognosis for Human CHS

Children wit h CHS may not manifest t heir suscept ibili ty to the disease for some yea rs, but once they do, the downhill course to death , though vari a bly prolonged, is usually inevi ta ble. Ironica l­ly, t hese children a re capable of making normal antibodies, developing competent delayed hyper­sensit ivity , and handling the usual childhood vira l infections.

The accelera ted phase of CHS is characteri zed by a .Jymphoreticul a r infil t rative process which responds to t he administrat ion of steroids ~nd antimitoti c chemotherapy. Dent et a l. (1966) fee l these characteristi cs warrant a pplying the term malignant to t he infiltra te . Ult imate ly t he bone marrow fa ils and the child dies, usually of a combination of overwhelming sepsis and a hemor­rhagic tendency related to the lack of neutrophil s and platelets . During the course, bes ides hepato­splenomega ly related to t he lymphoreticula r infil­trat e, there may be peripheral neuropathies, ap­pa rent ly related to t he in volvement of t he nerves wi t h either t he genet ic defect, t he infil t rates, or both (Lockm a n et a l. , 1967; Sung et a l. , 1969).

ANIMAL HOMOLOG UES AND GEN ETI CS

The a nima l models fo r CHS include t he Aleu­t ia n tra it in mink (Leader et al. , 1963) and a simila r mutan t in Hereford cattle (P adgett et al. , 1964) (Fig. 1) . A carefully documented mutant of C57Bl mice, known as beige, has a lso been de­scribed as morphologica lly homologous (Lutz ner et

530 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

a l. , 1967; Bennett et al. , 1969), and preliminary evidence suggests that t hese a nimals a lso show in creased susceptibili ty to infection if chal lenged in carefu lly controll ed experim ents (Padgett eta!. , 1970). Breeding experiments in a ll t hree an imal models have shown them to be t he results of the homozygous recess ive state of an autosoma l gene. All avai lab le evidence, inc ludi ng severa l reports of consa nguini ty (Kritzler eta!. , 1964) and one exten­sive family ped igree (Sadan et a l., 1965), indicates that the human condi tion is a lso an a utosoma l recess ive .

The animal homologues not only manifest vari­ous degrees of the clinical disease states but a lso offer the opportuni ty for epidemiologic study and controlled in vest igation by inoculat ion experi­ments. Padgett et a l. (1970) have reviewed and summarized t he resul ts of such stud ies a nd have stated that in large mink colonies and sma ll cattle herds the CHS animals show a n increased morbid­ity and mortality to many different bacteria and that ·the Aleutian mink are more susceptible to infection, morbidity, and death from t he Aleutian disease (AD) virus even t hough t he titer of AD virus in t he peripheral blood is not sign ifica ntl y different from that in other stra ins of mink. Pad­gett et a l. (1967) have a lso reviewed and compared the pathologic findings in the a nimal homologues with those in t he huma n condition (Table) .

GIGANT IS M Of CYTOPLAS MIC ORGANELLES

Their Nature and Distribution

From the t ime of the earliest reports, the mor­phologic abnormali ty of circulating leukocytes was suspected of being related to t he clin ica l difficul ­ties with infectious agents and with term inal marrow fa ilure experienced by t hese patients. Extens ive hi stochem ica l studies indicated that ab normally large granules contain essentia lly t he same materia ls as the normal gra nules cha racteris­tic of a non-CHS person . In an extensive ul tra­structura l in vestigation of the mink model , Lutzner et a !. (1965) demonstrated that giant granul es were found in practically a ll tissues that produce single-membrane-limited cytop lasm ic or­ga nelles. Miscellaneous studi es of huma n tissues corroborate t hi s findin g (Kritzler et a!., 1964; Windhorst et a l. , 1968a). Thus , it appears that t his single gene defect influences t he basic structure, though not necessa rily t he contents, of a wide variety of cytoplas mic granules t hroughout the body.

Origins

Davis et a l. (1971a, b) have used the electron microscope to study t he evolution of abnorma l gran ules in t he precursors and mature cells of mink

. bone marrow. They found that the neutrophil precursors of mink resemble t hose of rabb its in which at least two different types of gran ules a re t hought to arise from oppos ite sides of the Golgi

apparatus (Bainton and Farquhar, 1966, 1968a , b). In the Aleutian mink, the primary or azurophil granu les were abnorma l in size and structure in the early phases, and the giant forms of the mature neutrophils developed from t he fusion of these ab normal precursors. The seconda ry or "specific" gran ul es, on t he other hand, appeared to arise normally and to remain uninvolved in t he genera ­tion of giant granu les. T hus, on ly one of t h e subclasses of gran ules seen in norma l neutrophils (Baggiolini et al. , 1969) participated in t he genetic abnormality. Little is known about such subpopu­lations in any cells except polymorphonuclear leukocytes. As information about t he origins of other gra nules is reported , it shou ld be in teresting to see whether the CHS adds to our understandi ng of variations in cytoplasmic orga nelles. Davis eta!. (1971b) found what appear to be two subc lasses of granules in the mink basophils, but on ly one type, all abnorma l, in the eosi nophils. White (Windhorst et al. , 1968a), on the other hand , has photographs of human CHS eosinophils t hat suggest a vari ed gran ule constitution .

Attempts to Isolate Granules

Windhorst eta!. (1968a), using a sucrose density grad ient in a zonal ultracentrifuge, attempted to iso la te the gia nt gran ule fraction from the livers of Aleutian mink. The resu lts suggested that t he large gra nules are highly susceptib le to rupture by t he simplest of tissue homogenization procedures, which resulted in relatively increased amounts of soluble lysosoma l enzymes in the gradients from abnormal livers compared with those from non­Aleutian controls. Padgett 's group , using different techniques for ce ll rupture, were also unable to isolate enl arged CHS mink neutrophil granules (Padgett, G. A., unpub lished data).

Role of M embranes A major question in CHS is whether the giant

granules are formed by the failure of some primary mechanism that controls t he size of such organelles or by the fusion of gran ules that are of normal size when first formed. Neither possib ili ty necessari ly excl ud es the other, but because of the widespread nature of the defect a definite a nswer would have important implications: whichever process is at work, it is common to many if not a ll ce lls. Detection of a defect in t he membrane surrounding t he organelles cou ld provide a n a nswer t hat would supersede both the other a lternatives. Since mem­branes are composed of both lipid and protein , a defect in protein structu re wou ld be a direct reflection of t he mutant gene, whereas a defect in lipid composition would require a more complex ana lys is. Methodologies for investigating mem­branes are difficult, but some information about those of CHS is available .

The incorporation of P" in t he cel lula r phos­pholipid of resting and active phagocytes was norma l in CHS mink and cattle (Page and Pad-

CHEDIAK- HIGASHI SYNDROME 531

FIG. 1. A CHS Hereford ca lf with its mother to illustrate t he color dilution in t his stra in of bas ica lly red-ha ired anima ls. The dilu tion is assoc iated with susceptibility to disease, giant cytoplasmi c orga nell es , and early death. Transm iss ion is autosomal recessive.

TABLE

Comparative aspects of the Chediak - Higashi syndrome of man, mink , catt le, and mice

Cha racte ri sti c

Au toso ma lrecess ive mode of inh erita nce C linica l Studies

1. Inc reased susceptibi li ty to infec tion 2. Simi la r distribution of enl a rged granules 3 . Sim ila r morphology of enl a rged granul es 4. Sim ilar histochemistry of enlarged granules 5. Bleedi ng tendency 6 . Simi la r inl1a mmatory response 7 . Immunologic defic iency 8 . Pi gmentary dilution 9. Accelerated phase

10. Eleva ted muramidase levels Experimenta l Studies

1. Del ayed chemotax is in vitro 2 . Decreased serotonin levels 3 . Virus- like particles observed 4. Sequestration vacuoles observed 5. Abnorm a l serum lipid pattern 6. Autophagy 7. Abnormally frag ile enlarged granules 8. Abnormal bacte ri cidal ca pabili t ies of neutrophils 9. Abnormal membrane permeability

10 . Decreased renal tub ular catabolic function

gett, unpublished data); whereas Kanfer et a l. (1968) found an accelerated rate of phospholipid t urnover in the form of sphingomyelin in CH S patients. Windhorst (unpublished data) found

Man Mink Cattle Mice

yes yes yes yes

yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes yes ?

no no no ?

yes yes yes yes yes no no no yes no no no

yes yes ? yes yes yes yes yes occasiona l no occasiona l no occas iona l no no no yes no no no occasional no no no yes no no no ± ± no ?

occas iona l no no ? ? . yes ? yes

t ha t the incorporation of C" labeled inositol into t he phospholipids generated by enzymes from t he liver homogenates of Aleutian mink was norma l. H olland (1970) found no difference between nor-

532 THE JO URNAL OF INVESTIGATIVE DERMATOLOGY

mal ce ll s and t he red ce lls of t he mutant cattle in frag ili ty, tota l li pids, electrophoretic distribut ion of pho pholipids, a mino ac id compos it ion , and insolu ble prote in component .

Page et a l. (1962) found t hat blood 5-hydroxy­trypta mine levels were low in two patients, sugges­t ive of a defective plate let binding ca pac ity for serotonin . More recent observations by Holland in Padget t's group indi ca te t hat p late let serotonin is a lmost unmeasura ble in mutant mink, cattle , and mi ce and t hat added serotonin is not stored by the a bnorm a l platelets as effectively as by norma l ones . However , Davis et a l. (197lb) fo und t hat t he granul es of Aleut ia n mink platelets are morpholog­ically norm a l.

T HE PIGMENT AN OMA LY IN CHS

T he color defect seen in CHS affects the skin , ha ir, a nd eyes. A number of casua l descrip t ive terms have been applied to this phenomenon, e.g., partia l a lbini sm, oculocutaneous a lbinis m, a nd se mialbinism. However, since our information on t he basic mechanisms of pigmentation has become in creasingly more complete, our understanding of class ica l a lbinism has reached the molecula r level a nd we are forced to be more prec ise in our use of such terms. S uch preciseness is espec ia lly impor­ta nt since t hese te rms have rather spec ific implica­t ions in cl in ic eva luations.

The M elanosome as a Particular Cytoplasmic Organelle (reviewed in Fitzpatri ck et al. , 197la)

The ori gin of color in t he skin , ha ir , a nd eye of mammals derives principa lly from t he mela no­some, a spec ia l single membrane-bound cyto plas ­mic organell e produced by melanocytes. T he pig­ment it prod uces, melanin , is a highly insoluble, electron-dense hydroquinone ab le to absorb light across the ent ire vis ible spectrum wit h no charac ­terist ic absorpt ion peaks. It is la id down on the melanosome by t he act ion of a copper-conta ining enzy me, tyros inase, whi ch is apparent ly incor­porated into t he developing membra ne structure of the imm ature melanosome as it is fo rm ed in t he endoplas mic reticulum a nd t he Golgi a ppa ra tus (Se iji , 1967). Severa l subvari eties of mela nin a nd at least two ty pes of tyros inase , a ll under distinct gen etic cont rol, acco unt for t he different colors. In addi t ion, t he ava il a bili ty of t he substrate a mino acid , ty ros ine, to t his system may depend on genet ica lly determined ca rrier molecul es.

The color of t he epidermis and of t he ha ir shaft a rises from t he secretion of me lanosomes in to t he developing ke rat inocytes a nd ha ir matri x cells. Th e size a nd n umber of t he mela nosomes affect t he fin a l ha ir co lor; in addi t ion, t he shape may be refl ected in s ubtle vari at ions in color. T hese as­pects a re a lso under ge netic cont rol (Moyer, 1966) . Inhibi to rs of mela nin fo rmation in t he mela nocyte (Halprin and Ohkawa ra, 1967) a nd t he way in wh ich the keratinocyte does or does not "package"

t he melanosome play a role in the genetics of rac ia l color (Sza bo et a l. , 1969).

M any of t he resul ts upon which t hese statements a re made are based on studies of in bred mice, where at least 70 genes at 40 loc i influence t he color of the animal (Wolfe a nd Coleman, 1966) . Genera l­izing from such information to other ani ma l spe­cies is wa rranted by t he fact of homologous struc­tures a nd defects a mong ma mmals (e .g., ty rosinase defi ciency in t rue a lbini sm) as well as by the ancient nature of t he system, which is clea rl y present in birds, rept il es, amphibians, and fish as we ll as ma mmals (Bagna ra and Ferri s, 1971).

The CHS M elanosome

Since t he chemical nature of melani n is suc h that it is naturally electron-dense, ul trastructura l observat ions on t he pigmenta ry ab norm ali ty in CHS may be somew hat more s ignificant than those derived from cells whose organell es not only must be fi xed exte rna lly but a lso conta in high concen­t ra tions of lysoso mal enzy mes, e.g., neut rophils.

Windhorst et a l. (1966) have shown that the early melanosomes of human CHS are sometimes extremely la rge, even t hough t he la mellated mem­brane substructure may appea r to be qui te orderly (Fig. 2). In addi t ion, t he pattern of t he membranes of maturing CHS mela nosomes, ev idenced by in creasing melanization , suggests some fusion of smaller forms. On the basis of these findin gs and the demonstration of adequate tyrosinase act ivity in the epiderm al melanocytes of the pa ti ent, Wind­horst et a l. (1968b) proposed t hat CHS represents a human pigmentary dilu t ion based on a prima ry a lteration of t he structure of melanosomes. Wi th materi a l from t he sa me patient, Zeli ckson et a l. (1967) showed t hat t he destruction of t he melano­somes in the keratinocytes may be acclerated in CHS.

G iant melan in gra nules present in a nima l homo­log ues are a necessary cons iderat ion in defini ng CH S. In a se ri a l study of t he eye of t he developing feta l beige mouse, Lutz ner (1969) fo un d abnor­mally la rge mela nin gra nul es in both t he retina, whi ch conta in s mela nocytes deri ved from t he opt ic cup , (Moye r, 1961) a nd the choroid , which, like a ll cutaneous melanocytes, is neura l crest in origin . This di fference was evident in t he ea rli est fetuses in which pre mela nosomes could be detected . More­over, t hese ea rly giant granul e membra nes were disrupted and t he melanosome fibers were su r­rounded by vacuola r spaces . Both t he retina and t he choroid as we ll as t he pia a rac hnoid of human patients have been shown to conta in mela ni zed but giant mela nosome (Wind horst et a l. , 1968b; Be­doya et a!. , 1969).

The Pigmentary Dilution of CHS

When t he effects of t hese sub cellula r vari at ions a re observed in the in tact model, a part ia l dilution of t he bas ic color constit ut ion of the indi vidual

CHEDIAK- HIGASHI SYNDROME 533

Ftc. 2. Electron micrograph of melanocyte from ha ir follicle of CHS patient . Ea rly melanosomes of normal but giant substructure plus ev idence for fus ion of giant fo rms are presen t. (Osmium fixed, epon embedded, uranyl acetate sta ined, about x 31,000). (Reproduced from Windhorst, D. B. , Zelickson, A. S., and Good, R. A. 1966. Ched iak-Higashi syndrome: Heredi ta ry gigant ism of cytoplasmi c organelles. Science, 151:81- 83. Copyright 1966 by The American Assoc iat ion for the Advancement of Sc ience.}

res ul ts. This is part icul a rl y evident in t he mink in whi ch careful breeding acco mplishes exactly t hi s effect; the res ul t ing se ries of colors a re kn own as t he " blues" (S hackleford, 1950). Another i mpor­tant corolla ry is t hat a basica lly da rk individua l wi ll still be dark and his eye color will correspond ­ingly range from brown (in a child of dark-eyed heri tage) to t he tra nsl'ttcent "albino" eyes in a p erson whose "color base" is quite light. P hoto­phobia and nystagmus a re present even when the eyes show substant ia l co lor (Bedoya et a l. , 1969).

Clini cally, t he term pa rtia l a lbini sm has come to be associated wi t h pieba ldism , a genet ica lly deter­mined loca li zed absence of melanin -forming ce ll s equi va lent to spotting in mice. Ocu locuta neous a lbinism has come to mean t he lac k of melanin ­form ing capac ity with subdi visions relat ing to t he presence or absence of ty ros inase activity in t he m ela nocytes (F itzpatri ck and M ihm , 1971; Fi tzpa ­trick et al. , 197lb) . This ab norm a li ty is fu lly homologo us to t he a lbinism observed in lower forms , including mice a nd mink, wh ich in these spec ies is at a different genetic locus from t hat of C HS.

T hus, none of t he te rms com monly used to describe t he pigment in CHS convey t he fu ll implications a bout (1) t he genetic spec ificity of t he tra it, (2) its unique effect on t he structure of t he s ubcellular orga nell e carry ing t he pigment, (3) t he

part icul a r blu ish effect on t he observed color, someti mes even relat ive ly mild in degree, which this convey , or (4) t he specia l implications of th is particul ar melano omal a bnorma li ty on t he in ter­relat ionshi ps of pigment-forming ce ll s with other cells of t he body, pa rt icula rly t he circulat ing granulocytes .

DEFECTI VE DEFENSE MECHAN ISM S IN CI-IS

General Comments

In a ll the homologues studi ed , CHS correlates wi t h a lowered capacity to deal with various infectious age nts, including bacteria , viruses, a nd fungi (Candida alb icans) . This seems to be a nonspecifi c phenomenon in t he in tact host; t ha t is , where epidemiologic studi es have been made, Aleutian mink respond to a ll bacteria l a nd ome vira l age nts with increased morbidi ty a nd morta l­ity. On the other hand , t he poss ibili ty that a pa rti cul a r virus, e.g., infectious mononucleos is or viral hepat it is, is res ponsible for t he ultimate lymphoproli fe rative disease in t he children has not been rul ed out , and t hese syndromes a re men­t ioned in seve ral reports as being related to t he onset of t he accelerated phase of t he condi t ion. In additi on, t here is the unusual syndrome, Aleut ia n disease (AD) seen in Aleut ia n mink affected by a pa rt icul a r virus (Henson et a l. , 1966) .

534 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY

Studies on children (Page et al., 1962) and animals (Leader et a l. , 1963; Levine et a l. , 1966) have demonstrated a normal capacity to form antibodies and to develop delayed hypersensitiv­ity . These findin gs, combined with the morpho­logic abnorma li ties of granulocytes and wit h the development of new abilities to eva luate phago­cytes, have led to an intensive exam ination of the fun ctional capac ities of the neutrophil in CHS. It will be useful to report these studies in a way that will relate them to a systematic understand­ing of normal phagocyte fun ction .

Normal Phagocyte Functions and M ethods for Their Evaluation

Figure 3 (from Windhorst and Katz, 1972) is adapted from a more deta iled analys is of the basic methodology for studying phagocytes which has, in turn, been adapted for t he study ·of cli nical states (Windhorst, 1970) . The discovery and detailed ana lysis of a prototype defect in intracellular phagocytic function , chronic (fatal) granulomatous disease of childhood (CGD) (Good et a l. , 1968) have greatly st imulated interest in these importa nt components of innate defense.

Although Windhorst (1966), Windhorst et a l. (1968a), and Padgett (1967) were unable to detect a n a bnormality in huma n, mink, or cattle CHS leukocytes simila r to that in CGD, their results were preliminary, and as will be seen below, have been superseded. The recent work on CHS will be discussed according to the separate compart ments of Figure 3.

Maturation and Circulation of N eutrophils in CHS

Blume et a l. (1968) in a diffi cult study of CHS patients, found that hypersplenism is often present

©· • • ' ORDERLY ~ INTERACTION

BETWEEN PHAGOSOME and LYSOSOMES

All er Windi10I S! fldv ln l Med 1970

F 1G. 3. S implified di ag ram of the va rious parameLers of phagocyte function for whi ch methodologies a re ava il a­ble for study. The rap id evoluLion of t his type of work in cl inica l medicine relates to the discovery of chronic gra nulomatous disease of childhood , an X -linked defec t in the in t race llul a r events portrayed in the last two segments of t he diagra m. As indica ted in t he text, C HS has defects in various degrees in t he first two componen ts as well as the last two aspects of phagocyte fun ction. (F'rom Windhorst a nd Katz , 1972, by permiss ion).

when the chi ld is s ick, but if it is not, t he neutrophils have a norma l ha lf-life in the periph ­eral circulation. They also found increased levels of serum mura midase (lysozyme) and t he lack of a leukocytic response to etiocholanolone . These find­ings were taken to indicate a n increased destruc­t ion of the precursors of granulocytes within the marrow before they were released into the circula­tion. Padgett et al. (1970) found that cattle and mink do not have increased levels of serum mura­midase and t hat the levels of circu lat ing gra nulo­cytes in CHS and normal a nima ls were not differ­ent. In addition, Eklund et a l. (1968) found t hat leukocyte levels both in CHS mink with AD and in norma l mink with AD were normal.

Ka nfer et a l . (1968) interpreted t heir finding of increased leukocyte sphingolipid turnover as com­pati ble with increased lability of these cell s which a rapid bone marrow turnover would suggest. Both papers are compatible with White 's findin gs (1966, 1967) of increased cytop lasmic autophagy in human leukocytes and Windhorst et a l. 's (1968a) report of t he proba ble lability of mink li ve r lyso­somes in CHS .

Chemotactic Responses

Despite an early report by Page et a l. (1962) that in vivo skin-window studies do not revea l abnorma l cell migrat ion (Windhorst et a l. , 1968a) , Clark and Kimball (1970) a nd Clark et a l. (1971) have found that when ex posed to s tandard chemotactic stim­uli , the granulocytes of huma n a nd mink CHS are defini t ively ab normal in their capacity to migrate in vitro in a Boyden chamber. There was no defect in the a bility of CHS serum to generate chemotac­tic factors.

Surfac e Intera ction , Engulfment, and Degranulation

The reports of Page et a l. (1962) and Saraiva et a l. (1959) on opsonic indices, as well as t he quantitative bactericida l studies reported below, indicate that the uptake of bacteria by CHS leukocytes is not defect ive when adequate amounts of normal or CHS serum a re present. Padgett (1967) and Root et al. (1968) have found that t he abnorm a lly la rge PMN granu les fai l to fu se wit h phagosomes whereas norma l granules function properly. Padgett et a l. (1970) suggest that t his is ev idence for an ab norma l stability of t he gian t granular membra ne ra t her than a n increased lab il ­ity suggested by White (1967).

Intracellular Bacterial K illing and Degradation

Root (1971), Stossal eta!. (1972) , a nd Root et a l. (1972) have studied hum an CHS extensively for phagocytic uptake a nd other functio ns, ut ilizing va rious organ isms in the qua nt itative bactericida l tests and a na lyz ing metaboli c activity as well. They find that the kinet ics of t he abi lity of CHS phagocytes to kill organisms is such t hat a defect can be detected for E. coli and C. albicans on ly at a

CHEDIAK- HIGASHI SYNDROME 535

time interva l of 20 minutes, whereas t he a bnormal killin g of Staph. aureus, D-st reptococc us, a nd a rough strain of pneumococcus can be seen up to two hours, but not later. They in te rpreted t heir results to indicate that t he ea rly defect in bacteria l killing is a consequence of the s low-to-a bsent fus ion of the peroxidase-contain ing giant granul es with t he phagosome a nd raised the possibility t ha t t his fu sion is important in the ea rly kinetics of normal phagocy tic killing. Davis (1970) found C HS cattle leukocytes deficient in bac teri cidal ca pacity.

In t his same genera l category, though not stri ctly involving the phagocytic system assoc iated with defense aga inst infections, Prieur a nd Padgett 0970) a nd Prieur et a l. (1972) have s hown that t he Proxima l co nvoluted rena l tubu les of CHS mink and mi ce handle a n in t rape ri tonea l dose of horse­radish peroxidase differently from and more slowly tha n norma l mink kidneys . They suggest that an understa nding of t his finding will contribute to our knowl edge of the genes is of the lupus ery­thematosus- like glomerul onep hri tis seen in Aleu­tian di sease. In addit ion, it may correlate ve ry well with the delayed fusion of gran ules t hat has been obse rved in the neutrophil.

CONCL US ION

The C hediak-Higashi sy ndrome see ms to repre­sen t a defect in subce llul a r structu re whi ch in turn is refl ected in function a l abnorma li t ies of seem­in gly diverse cell lines. This correlation is , of cou rse, not surpris ing in view of t he many meta­bolic features s ha red by a ll ma mma li a n cells. However, the s ingle membrane-bound cytop lasmic organel les can legit imately be regarded as distinc­tive for the cell in which t hey occur, that is , t hey express specifically the full differentiative ca paci ty of a ny give n ce ll , be it neutrophil or melanocyte. In this sense, such structures cou ld be ex pected to express the unique cha rac teristics of the differen­t iation of a given cell. The Chediak-Higashi sy n­drome indicates that not a ll t he characteristics of such gra nules are unique , but at least one feature under genetic control is common to most if not all such organe lles.

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