Acta Troptca, 46(1989)249-256 249 Elsevaer

ATP 00033

Hypogonadism and ecdysteroid production in Loa loa and Mansonella perstans filariasis

J. Lansoud-Soukate 1, A. Dupont 1, M.L. De Reggl 2, G E. Roelants 1 and A. Capron 3

1International Centre for Medz~al Research ( CIRMF), Francevdle, Gabon, 2Irnmunotech, S A , Marsedle, France, and 3Centre d'lmmunologae et de Blologw Parasltatre, umtb mt',te 1NSERM, Instttut

Pasteur, Ldle, France

Possable endocrlnologacal repercussions of mfecnon with Loa loa and Mansonella perstans filanae were studaed in Gabonese subjects Mlcrofilaremlc males were compared wath amacrofilaremlc controls In the infected group 13/105 subjects (12%) presented only abnormally low serum levels of testosterone ( < 4 ng/ml), 25/105 (24%) only abnormally high serum levels of gonadotrophms, FSH (> 15 mIU/ml) and LH ( > 20 mIU/ml), and 22/105 (21%) presented anomahes m both testosterone and gonadotrophm levels One out of 68 control subjects had 3 6 ng/ml serlc testosterone and all had normal levels of gonado- trophms Ecdysteroads were detected ( > 0 025 ng/ml) m the serum of 87/97 (90%) m~crofilaremlc subjects (GM 0 123 ng/ml) compared to 12/64 (19%) controls (GM 0 030 ng/ml) Ecdysteroads were detected m the urine of all subjects, infected (GM 8 468 ng/ml) as well as control (GM 1 245 ng/ml) The hormonal perturbations were correlated wath the levels of Loa loa macrofilaremla but not wath those of serum and urinary ecdysterolds These results demonstrate that macrofilaremlc subjects often show endocnnal signs of hypogonadasm and present apprecmble levels of ecdysteroads m serum and urine A darect role for parasitic ecdysteroads in hypogonadasm remains to be demonstrated

Key words Fdarlasls, Loa loa, Mansonella peratans, Hypogonadasm, Ecdysteroads

Introduction

It is estimated that 300 mflhon people in the world are infected wtth filarlae (Faro, 1981a) Loa loa filarlasts affects 2 to 3 milhon subjects m Central Africa (Faro, 1981 b) and Mansonella perstans filanasts 40 to 80% of mdlvtduals hvmg m Central and Western Afraca (Hawking, 1977) The geographical dlstrtbution of loalasts is coincident with a zone of hypofecundlty with btrth rates of only 2 0 to 3 5%, whereas surroundmg regions have rates of about 6 0% (INSEE, 1960-61, Romantuk, 1968, Meheus et a l , 1986) Certam filarlases such as those caused by Wuchererla bancroJtt (Ltchtenberg and Medma, 1957, Iturregut-Pagnan et a l , 1976) and, to a lesser extent, Onchocerca volvulus (Strangway and Strangway, 1951, Nelson, 1958, Gratama, 1969), can reduce lesxons of the male gemtal tract That the same is true for L loa and M perstans infections is not clear (Gordon et a l , 1950, Langutllat et a l , 1978a, b) To our knowledge, the endocrine repercussions m the host of filarla infectxons have

Correspondence address Dr J Lansoud-Soukate, Centre Internanonal de Recherches M6dlcales de Francevllle, B P 769, Francevflle, Republlque Gabonalse

0001-706X/89/$03 50 c~ 1989 Elsevier Science Pubhshers B V (Biomedical Division)

250

however not been investigated Thus we have looked for endocrine abnormahtles associated with L loa and M perstans mlcrofilaremla in naturally infected subjects Ecdysterolds are hormones controlling essential events of development and repro- ductIon m arthropodes and probably other protostomlan phylae and have been detected in helminths including filariae (Koolman, 1984, Rees and Men&s, 1984) In certain cases they have been shown in the biological fluids of parasitized vertebrates (Koolman, 1984, Nlrd6 et a l , 1984a,b) The possible correlation of parasite ecdysterolds and endocrine repercussions of L loa and M petstans' refections is investigated

Materials and Methods

We tested 776 men aged 20 60 years from the province of Haut-Ogoou6, Southeastern Gabon, for the presence of L loa and/or M perstans mlcrofilaremla Of 367 mlcrofilaremlc subjects, 262 were rejected for presenting other parasites in blood and/or stool, of 409 amlcrofilaremac subjects, 341 were rejected for the same reason Mlcrofilarlae detected, using the Knot t ' s concentrat ion method (Markell and Voge, 1981), were distributed as follows m the blood of the remaining 105 mlcrofilaremlc subjects 18 L loa only, 48 M perstans only and 39 with both None of these mlcrofilaremlc subjects had obvious signs of hypogonadlsm on physical examination in the field Controls consisted of the remaining 68 amlcrofilaremxc subjects (hmlt of detection 1 organlsm/ml of blood) without clinical signs or history of filanasls No other parasites were detected in the blood or stool of the 173 subjects selected for this study All samples were collected between 10 00 h and 15 00 h All subjects were of comparable SOClO-economical status and no signs of malnutri t ion were detected

Serum testosterone was measured for all subjects by the competitive radiolmmu- noassay of Abraham et al (1977) (using T R K 402 Amersham-France reagents) The results are expressed as ng/ml, the limit of detection was 0 1 ng/ml and normal values were considered to be 4 to 8 ng/ml

Serum levels of folhcle-stlmulating hormone (FSH) and lutelnizlng-hormone (LH) were measured by radloimmunoassay using commercial kits (CEA-SORIN, Ons- Industries, St Quentin en Yvellnes, France) The results are expressed as mIU/ml, the limit of detection was 1 5 mIU/ml and normal values were considered to be 5 to 15 mIU/ml for FSH and 4 to 20 mIU/ml for LH

Quantity of material available allowed ecdysterolds extraction from the serum and urine of only 97 mlcrofilaremlc and 64 control subjects Ecdysterolds were measured according to the method of De Reggl et al (1975) Results are expressed as ng 20- hydroxy-ecdysone equivalent per ml, the limit of detection was 0 025 ng/ml

Levels of ecdysteroids, testosterone, FSH and LH were log-normally distributed, with the exception of ecdysterolds in urine, and central tendencies were measured using the geometric mean The slgmficance of group differences was analysed by the Student's t-test, the Behrens-Fisher's test, the F-test and/or the ~2 test with Yate 's correction (Armltage, 1971) The significance of group differences for ecdysterolds In urine, which were not normally distributed, was analysed by the Kruskal-Walhs ' H- test (Siegel, 1956) Regression curves of correlations were calculated and eventually produced (Fig 2) with the Microsoft Chart R on a Macintosh R computer

Results

251

The presence of mlcrofilarlae in blood was independent of subject age in the present study L loa mlcrofilanae ranged from 1-86 200/ml (median 540/ml) and M perstans from 1-4 300/ml (median 100/ml) Hormonal levels in all groups described below were also independent of subject age

In the mlcrofilaremlc group, 13/105 subjects (12%) presented only abnormally low levels of serum testosterone (< 4 ng/ml), 25/105 (24%) only abnormally high levels of serum gonadotrophms FSH (>15mIU/ml) and LH (>20mIU/ml), and 22/105 (21%) presented both low testosterone and high gonadotrophm levels Only one out of 68 control subjects had borderhne (3 6 ng/ml) serum testosterone and all had normal levels of gonadotrophms Means are presented in Fig 1 Mean hormonal

m [U/m I ng/ml

14

12

10

0 ~

FSH LH Testosterone

Fig l Geometric m e a n + s t a n d a r d error of the mean of gonadot rophms FSH and LH (mlU/ml) and testosterone (ng/ml) m mlcrofilaremlc ( • ) and amlcrofilaremlc ([]) subjects

252

levels did not differ for the three types of mlcrofilaremla L loa alone, M perstans alone and both together However, the 18 subejcts presenting only L loa mlcrofilar- lae showed a negative correlation between the levels of mlcrofilaremla and the serum testosterone (r = - 0 64, p < 0 005) and/or a positive correlation between the levels of mlcrofilaremla and FSH levels ( r - 0 71, p < 0 001) (Fig 2) These correlations were not found m subjects presenting pure M perstans mlcrofilaremla and there was no correlataon between L loa mlcrofilaremla and LH levels

Table 1 shows the levels of ecdysterolds measured in serum and urine Serum ecdysterolds were detected m 87 of the 97 mlcrofilaremlc subjects for which sera were

TESTOSTERONE (ng/ml)

10

It

8

4

2

0

0

I I I I

100 1000 10000 100000

Loa Ioa rnJcrofilaremla (/ml)

253

available and also m 12 of the 64 controls tested, albert m lower concentrations The difference between the means for the two groups was highly significant (p < 0 001) Urinary ecdysterolds were detected in all subjects although again m very significantly lower mean concentration in the control than m microfilaremic subjects (p < 0 001) There was no correlation between serum and urinary levels of ecdysteroids nor between the species and degrees of mlcrofilaremla and ecdysteroid levels m either serum or urine

Host hormone levels were not correlated with ecdysterotd levels in etther blood or

FSH (mIU/ml)

80

70

60

50

40

30

20

10

B

o I I I I

10 100 1000 10000 100000

Loa Ioa mmrofilaremta (/rnl)

Fig 2 Regression curve of correlatmn between testosterone (A) or FSH (B) serum and Loa loa m~crofilaremm

254

TABLE 1

Ecdysterolds m subjects bearing microfilarlae and controls

Ecdysterolds (ng/ml)

Sample Subjects n Geometric mean Range

Serum mlcrofilaremlcs 97 0 123 a < 0 025-3 605 controls 64 0 030 < 0 025 0 485

Urine mlcrofilaremlcs 97 8 468 b 0 275 87 215 controls 64 1 245 0 255-44 045

aBehrens-Flsher's test d= 10 9, q=29 01, p <0 001 bKruskal-Walhs's test H-46 5, p <0 001

urine Indeed, o f the 10 mlcrof i laremlc subjects wi thou t de tec table ecdys tero lds in serum, 5 had h o r m o n a l a b n o r m a h t i e s

Discussion

The levels o f microf i la remla found in the subjects s tudied here covered the range repor ted m the l i te ra ture (Fain , 1981, Hawking , 1977) Such h o r m o n a l d i s turbances as those detected in 60/105 (57%) microf i laremlc subjects in our s tudy could p lay a role in hypofecund i ty in Cent ra l Africa. Paral le l decreases o f tes tos terone levels and increases o f g o n a d o t r o p h l n levels might indicate a p r ima ry tes t lcular defect (Huang et a l , 1984, Ib rah lm et a l , 1983) Decrease in tes tos terone level wi thout e levat ion o f g o n a d o t r o p h l n s , observed in 13/105 (12%) o f mlcrof i laremlc individuals , could also po in t out to a supra-test~cular mechan i sm Tha t tes tos terone could be efficiently m e t a b o h z e d by paras i tes has yet to be excluded Ecdys te roIds were found in serum and urine o f m~crofilaremic indiv iduals as has been the case in o ther paraslUzed ver tebra tes ( K o o l m a n et a l , 1984, Nlrd6 et a l , 1984a,b) Ecdys te ro lds were also found m low concen t ra t ions in 19% amicrof i la remic cont ro ls This could be due to some subjects car ry ing adu l t worms wi thout mlcrof i lar la , as has been demons t r a t ed for 22 out o f 33 subjects present ing with a L loa eye passage ( D u p o n t et a l , 1988) Moreove r , some o ther he lmln th paras i te infect ions could have escaped detec t ion F u r t h e r m o r e , h u m a n food conta ins phy toecdysones which are ra ther widespread an p lan ts Be tha t as it may, ecdys te ro ld levels were significantly higher m the mJcrofilarem~c than in the amicrof i la remic g roup

H o r m o n a l p e r t u r b a t i o n seems however cor re la ted with the degree o f mlcrofi lare- mxa and not with ecdys tero id levels Whi le ecdys tero lds could p lay a role m h y p o g o n a d l s m , e g t h rough their ac t ion on cyclic A M P metabo l i sm (Cat t et a l , 1979, 1980, Ca ta l an et a l , 1979), a role for these paras i t ic ho rmones in this p h e n o m e n o n remains to be d e m o n s t r a t e d

Acknowledgments

C I R M F is 70% financed by the Sta te o f G a b o n and 30% by funds p rov ided by Elf- G a b o n We thank the endocr ino logy l a b o r a t o r y o f C I R M F for measur ing testos-

255

terone, FSH and LH, Mrs L Trotteln and Mr E Ruelle for excellent assistance in computer analysis, Drs C Chlzzollm, R W Cooper, T E Egwang, D Mavoungou, and M Plnder for rewewlng the manuscript and Mrs M A Molto for expert secretarial assistance

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