Vol. 13, No. 1JOURNAL OF CLINICAL MICROBIOLOGY, Jan. 1981, p. 30-350095-1137/81/010030-06$02.00/0

Characterization of Antibody Activity in OligoclonalImmunoglobulin G Synthesized Within the Central Nervous

System in a Patient with Tuberculous MeningitisJORGEN KINNMAN,' HANS LINK,'* AND ARIL FRYDÉN2

Departments of Neurology' and Infectious Diseases72 University Hospital, S-581 85 Linkôping, Sweden

Thin-layer polyacrylamide gel isoelectric focusing of cerebrospinal fluid (CSF)and serum obtained from one patient 48 and 65 days after the onset of tuberculousmeningitis revealed five oligoclonal immunoglobulin zones in CSF without anycounterpart in serum, indicating local immunoglobulin production. Subsequentimmunofixation with specific antisera revealed that three of the zones consistedof immunoglobulin G lambda, and two consisted of immunoglobulin G kappa andimmunoglobulin G lambda present simultaneously. Immunofixation with Myco-bacterium tuberculosis and bacillus Calmette-Guérin (BCG) as antigens andautoradiography revealed zones of specific antibodies in the CSF which, regardingmobility, corresponded to oligoclonal and polyclonal CSF immunoglobulin Gzones. No antibody activity was detectable in the corresponding serum, indicatingthat the antibodies present in CSF were synthesized within the central nervoussystem. In seven control patients (three with multiple sclerosis, four with chronicinflammatory central nervous system diseases of unknown cause) with oligoclonalCSF immunoglobulin, no evidence for local production of antibodies against M.tuberculosis or BCG was detectable. Immunofixation with M. tuberculosis orBCG as antigens and autoradiography may prove to be a useful diagnosticcomplement to conventional techniques in patients with suspected tuberculousmeningitis.

Although cell-mediated immunocompetenceis of the utmost importance for the course andoutcome of human tuberculous infections, thehumoral immune response might also play animportant role. In tuberculous meningitis (TM),an intensive humoral immune response withinthe central nervous system (CNS) and charac-terized by the intrathecal synthesis of oligoclonalimmunoglobulin may occur (2; J. Kinnman, A.Frydén, S. Eriksson, E. Moller, and H. Link,Scand. J. Immunol., in press). The phenomenonof local immunoglobulin synthesis is character-istic of a great variety of inflammatory disordersaffecting the CNS (for a review, see reference 5).The antibody character of locally produced oli-goclonal immunoglobulin G (IgG) has been de-fined in some of these disorders, e.g., in subacutesclerosing panencephalitis (8, 13), whereas inothers, such as multiple sclerosis, the specificityof oligoclonal IgG synthesized within the CNShas been only partly clarified (R. Rostrom, H.Link, M. A. Laurenzi, S. Kam-Hansen, E.Norrby, and B. Wahren, Ann. Neurol., in press).

Characterization of locally produced immu-noglobulin in etiologically well defied inflam-matory CNS diseases, such as TM, is of impor-tance for two reasons: (i) identification of the

antibody detectable in locally produced immu-noglobulin should be evaluated as a diagnostictool and, therefore, its specificity and its appear-ance as well as disappearance during the courseof disease should be established and (ii) carefulanalysis of, inter alia, the temporal profile of theimmunoglobulin synthesized within the CNS re-garding amount, class, and antibody specificitymay increase our understanding of the localimmune response in disorders of hitherto un-known pathogenesis, such as multiple sclerosis.

In this paper, oligoclonal immunoglobulin pro-duced within the CNS in one patient with TMis characterized regarding class, light chain type,and antibody specificity. Using antigen immu-nofixation and autoradiography, it is shown thatsome of the oligoclonal IgG zones detectable inthe patient's cerebrospinal fluid (CSF) by thin-layer polyacrylamide gel (PAG) isoelectric fo-cusing (IEF) contain antibodies directed againstMycobacterium tuberculosis and against bacil-lus Calmette-Guérin (BCG).

MATERIALS AND METHODSSubjects. (i) Patient with TM. A previously

healthy 33-year-old woman contracted signs andsymptoms suspected to be due to TM on 21 March

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OLIGOCLONAL IgG IN TUBERCULOUS MENINGITIS 31

1976. The diagnosis was confirmed by identification ofM. tuberculosis by culture of three consecutive CSFspecimens taken on 4, 9, and 20 April. Direct micros-copy did not reveal M. tuberculosis in the CSF. Thepatient's condition improved after treatment with tu-berculostats instituted on 11 April. Steroids were notgiven. She was discharged completely recovered 3months later. Lumbar punctures were carried out onseveral occasions after admission, and a serum speci-men was always taken simultaneously (see Table 1).

(Hi) Control patients. CSF and serum for antigenimmunofixation were also obtained from three pa-tients with clinically definite multiple sclerosis andfour patients with chronic meningoencephalitis of un-known etiology. Repetitive CSF cultures for M. tuber-culosis in the four latter patients were negative. Allseven patients were ambulatory. They displayed oli-goclonal immunoglobulin on routine agarose gel elec-trophoresis of CSF.Routine CSF and serum studies. After cell

counting by phase-contrast microscopy, the CSF wascentrifuged. Albumin, IgG, and IgA were determinedsimultaneously in the CSF and serum, as were theCSF/serum albumin ratio as a variable for the blood-brain barrier function and the CSF IgG index [i.e., theratio (CSF IgG/serum IgG):(CSF albumin/serum al-bumin)] and the corresponding CSF IgA index asvariables for the demonstration of intrathecal synthe-sis of IgG and IgA, respectively. The age-dependentnormal values previously described for the CSF/serumalbumin ratio were used (11). The upper normal valuesfor the CSF IgG index and the CSF IgA index in ourlaboratory are 0.7 and 0.6, respectively. Elevated val-ues indicate synthesis within the CNS of the immu-noglobulin in question (9). The IgG synthesis ratewithin the CNS was also calculated (12). Our uppernormal value is 10 mg/24 h.

Thin-layer PAG IEF. IEF was carried out onAmpholine-prepared thin-layer PAGs (PAG plates;LKB, Stockholm, Sweden), pH range 3.5 to 9.5, aspreviously described (3). In short, CSF concentratedby ultrafiltration at 4°C in collodium bags (Membran-filter; Sartorius, Gottingen, Federal Republic of Ger-many) to an IgG level of about 3 g/liter and serumdiluted with 0.05 M NaCl to the same IgG level wereapplied to the gel at a pH between 6.2 and 6.4 atamounts of 10 Pl, corresponding to about 30 gg of IgG.On every plate pooled blood donor serum diluted with0.05 M NaCl to an IgG level of about 3 g/liter was runas a reference, and carboxy-hemoglobin was run as amarker. The protein patterns obtained on PAG IEFofCSF were compared with those ofthe correspondingserum and of normal serum. To define the results, thepH range was divided into three regions: I, pH 3.5 to6.4; II, pH 6.5 to 8.6; III, pH 8.7 to 9.5. Zones found inCSF in pH regions II and III in addition to those seenon PAG IEF ofthe corresponding ser run in parallelwere considered to represent oligoclonal immunoglob-ulin (3, 4a).The investigations carried out on CSF and serum

after PAG IEF are summarized in Fig. 1.Immunofixation with specific antisera. For the

characterization of immunoglobulin bands (regardingclass and light chain type) found on PAG IEF, im-

Antiserum immunofixationfor definition of clasm andlight chain type of immu-noglobulin zones

Antigen inmmunofixation andautoradiography for detec-tion of antibodies in im-munoglobulin zones

PAG IEF carried out on CSF and serum obtainedsimultaneously

1Incubation with cellulose

acetate strip dipped inmonospecific antiserum

Specific antigen-antibodycomplexes are formed inthe PAG plate

tStaining for detection of

zones of antigen-antibodycomplexes in the PAGplate

Incubation with antigen-con-taining agarose gel plates

Specific antigen-antibodycomplexes are formed inthe antigen-containingagarose gel plate

1~Autoradiography for detec-

tion of antibodies com-plexed in the agarose gelplate

Comparison of zone patterns with those from reference runof CSF and serum

FIG. 1. Scheme ofprocedures used for classifica-tion ofoligoclonal immunoglobulin zones demonstra-ble by thin-layer PAG IEF of CSF and serum from apatient with TM.

munofixation was done after the run, using monospe-cific antisera (Dakopatts, Copenhagen, Denmark)against y chains, a chains, and kappa and lambdaBence-Jones proteins (4, 4a).Antigen immunofixation and autoradiogra-

phy. (i) M. tuberculosis and BCG. M. tuberculosisstrain 37 (kindly supplied by B. Wahren, NationalBacteriological Laboratory, Stockholm, Sweden), in-activated by heating to 100°C and freeze-dried, wasdiluted in 0.05M NaCl. BCG (National BacteriologicalLaboratory), freeze-dried, was similarly diluted. Beforeuse in the antigen immunofixation experiments, thepreparations of M. tuberculosis and BCG were soni-cated in the cold with a 150-W Ultrasonic Disintegra-tor (mark 2; MSE, Crawley, England) with a 3-mmexponential probe at an amplitude of 12 to 14 ,m forsix 10-s bursts and centrifuged at 3,000 x g for 10 minat room temperature. The supernatants thus obtainedwere used as antigens in antigen immunofixation.Three different concentrations of the antigens wereused, namely, 400, 50, and 5 pl, respectively, of M.tuberculosis per 5 ml of agarose gel, and 500, 50, and5 id, respectively, of BCG per 5 ml of agarose gel.

(ii) Viral antigens. Ten percent (vol/vol) suspen-sions of the Lec strain of measles virus cultivated inVero cells, complement fixation titer of 160 (kindlysupplied by E. Norrby, Karolinska Institute, Stock-holm, Sweden); herpes simplex virus (HSV) type 1cultivated in baby hamster kidney cells, complementfixation titer of 320 (E. Norrby); concentrated extra-cellular mumps virus propagated in chorioallantoicmembrane cells, complement fixation titer of 160 (D-449; Orion, Helsinki, Finland); and Mycoplasmapneu-

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32 KINNMAN, LINK, AND FRYDÉN

moniae, complement titer of 32 (D-433, Orion), werekept at -20°C until used as reference antigens in theantigen immunofixation experiments. Before use, theviral antigen preparations were sonicated as describedabove for three 10-s bursts and centrifuged for 10 minat 1,500 x g at room temperature. The supernatantswere used as antigens in antigen immunofixation. Thefinal concentrations per 5 ml of agarose gel were 150M1 for measles virus preparation, 10 Ml for HSV, 50 tlfor mumps virus, and 200 1l for M. pneumoniae. Theconcentration of each type of antigen per 5 ml of gelwas determined in calibration experiments.

(iii) Detection of antibodies by antigen im-munofixation and autoradiography. CSF and thecorresponding serum were run in duplicate on PAGIEF, one specimen being used for detection of anti-bodies in immunoglobulin zones, the other as refer-ence. The presence of specific antibodies was detectedby antigen immunofixation followed by incubationwith '25I-labeled (code IMS 30; Radiochemical Centre,Amersham, England) rabbit anti-human immunoglob-ulin antibodies (code A-107, Dakopatts) and autora-diography as described elsewhere in detail (10; Ros-trom et al., Ann Neurol., in press). In short, the PAGIEF plate to be used for antibody detection wasquickly washed with phosphate-buffered saline, pH7.2 to 7.4, immediately after IEF. The antigen-contain-ing agarose gel plates were placed with the gel in directcontact with the PAG IEF plate, with a thin layer ofphosphate-buffered saline in between. Only the pHregion 6.5 to 9.5, i.e., pH regions II and III, was covered.PAG IEF and antigen-containing agarose gel plateswere kept together for 5 to 8 min, during which timeantibodies moved to the agarose gel plates to formantigen-antibody complexes. Up to five different an-tigen plates were successively placed on the same PAGIEF plate. After washing the plates in phosphate-buffered saline overnight and repeatedly pressingthem under filter paper (10 min) and washing them inPBS (15 min), the plates were incubated with 1251_labeled rabbit anti-human immunoglobulin antibodyfor 12 h. The plates were then washed in phosphate-buffered saline, pressed under filter paper three times,and, finally, washed in distiled water, pressed, and airdried. The antibodies precipitated on the antigen-con-taining agarose gel plates were indicated by autora-diography carried out for 6 to 36 h with Kodak X-Omat L. To avoid artefacts on autoradiography, thefirst plate to be used for antigen immunofixation con-tained only agarose (agarose control plate). Some au-toradiograms obtained from such plates revealed faintzones mostly in the vicinity of the application slit,which corresponded to protein zones seen on PAGIEF. The occurrence of such artefacts was efficientlyprevented by a quick phosphate-buffered saline washof the PAG IEF plates immediately after the run andbefore subsequent antigen immunofixation.The zone patterns obtained at autoradiography

were compared with those from PAG IEF of thecorresponding CSF and serum run in parallel as ref-erences and also with those obtained after Coomassieblue staining of the PAG IEF plate after it had beenused for antigen immunofixation. Intrathecal antibodysynthesis was concluded to occur when (i) one or more

of the zones on autoradiography were observed in CSFonly but not in the patient's serum and (ii) one ormore of the zones on autoradiography were distinctlymore pronounced in CSF than in the patient's serum.

RESULTSThe patient with TM displayed throughout

the observation period-even at the time whenshe had recovered clinically-evidence of in-trathecal synthesis of IgG as reflected by ele-vated CSF IgG index values and increased IgGsynthesis rate per 24 h (Table 1). The CSF IgAindex was elevated between days 48 and 97,indicating IgA synthesis within the CNS. Onroutine agarose gel electrophoresis, two oligo-clonal immunoglobulin bands were found in allfour CSF specimens, but not in the correspond-ing sera.Immunoglobulin class and light chain

type of oligoclonal zones found in CSF inthe patient with TM. PAG IEF of CSF andserum from the patient 48 and 65 days after theonset of symptoms due to TM revealed fivezones in CSF in pH region III, i.e, pH 8.7 to 9.5,without any counterpart in the correspondingserum, reflecting the presence of oligoclonal im-munoglobulin. Immunofixation with monospe-cific antisera revealed that all five zones con-sisted of IgG, three of them with light chains ofthe lambda type and the remaining two withlight chains of types kappa and lambda presentsimultaneously (Fig. 2).Demonstration of antibody synthesis

within the CNS. Antigen immunofixation ofCSF and serum obtained on days 48 and 65 afterthe onset ofTM (Table 1), using M. tuberculosisas the antigen, revealed on the autoradiogramfive zones in the CSF specimen from day 48 andthree zones in the CSF specimen from day 65,but no zones in the corresponding serum (Fig.3). These zones thus represented antibodiesagainst M. tuberculosis synthesized within theCNS. Similar zone patterns present only in CSFwere also obtained when BCG was used as theantigen.Two of the zones of antibodies against M.

tuberculosis and BCG found on the autoradi-ograms of CSF corresponded regarding mobilityto two of the five zones identified as oligoclonalIgG. The remaining M. tuberculosis-specific an-tibody zones from the autoradiograms corre-sponded to polyclonal IgG zones.With HSV, measles virus, mumps virus, and

M. pneumoniae as reference antigens, the onlyabnormality found was one zone of HSV anti-bodies in CSF and serum obtained on day 48and two zones of HSV antibodies in CSF andserum obtained on day 65. These zones were

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OLIGOCLONAL IgG IN TUBERCULOUS MENINGITIS 33

TABLE 1. CSF findings in a patient with TM

Days after onset 10o Leukocytes/liter CSF/serum ai- IgG synthesis d CSF/serum glu-of disease (p1morponuc1ears fbumin ratio' CFIGidxrate/24 h (mg)' CSF IgA index oerto

14 102 (14/88) 26.0 0.8 51 NDf 0.248 58 (1/57) 14.7 1.2 54 1.1 0.265 38 (0/38) 16.0 1.1 43 1.1 0.497 13 (0/13) 9.5 1.0 19 0.9 ND

a Normal value, '6.8.b Normal value, sO.7.C Normal value, <10 mg.d Normal value, -0.6.' Normal value, >0.5.fND, Not done.

pH

4.0-

6.5-

8.6-

A B C DFIG. 2. Patterns from thin-layer PAG IEF and

subsequent immunofixation of CSF (A through C)and serum (D) from a patient with TM meningitis,using antisera against y chain (A and D) and Bence-Jones proteins type kappa (B) and type lambda (C).Arrows denote oligoclonal IgG zones.

much stronger in CSF than in serum, indicatingintrathecal synthesis. The mobilities on the au-

toradiograms of the HSV-specific zones differedfrom those of the M. tuberculosis- and BCG-specific zones.The zones of HSV antibodies had their coun-

terparts in polyclonal IgG zones in CSF andserum.

All runs were repeated on another occasion,using the same antigens, with the same results.For reference, CSF and the corresponding se-

rum from each of seven patients with otherneurological disorders and oligoclonal immuno-globulin in CSF were also investigated by im-munofixation with M. tuberculosis and BCG asantigens and subsequent autoradiography. Nozones of intrathecally synthesized antibodiesagainst either M. tuberculosis or BCG were de-monstrable on the autoradiograms.

DISCUSSIONTM is a rare disease in Scandinavia, and

therefore material from only one patient wasavailable for this investigation. From previousstudies it is known that oligoclonal immunoglob-ulin bands may appear in CSF in TM alreadyduring the first days of the disease (E. C. Laterre,personal communication). However, oligoclonalimmunoglobulin is probably not found in CSFin every patient with TM. We have found thisin 1 out of 2 patients (Kinnman et al., in press),and Laterre has found this in 5 out of 13 patients(2).Immunofixation with monospecific antisera

revealed a predominance in number of oligo-clonal IgG lambda bands in the CSF from ourpatient with TM. A predominance of IgGlambda has previously been demonstrated inacute aseptic meningitis with oligoclonal bandsin CSF (1, 7), whereas a predominance of IgGkappa has been registered in multiple sclerosis(7). It is therefore possible that the oligoclonalimmune response within the CNS, with respectto intrathecal production of predominantly IgGkappa or IgG lambda, differs in various inflam-matory CNS diseases.Two of the five oligoclonal IgG zones demon-

strable on PAG IEF of CSF obtained on twodifferent occasions from our patient with TMcontained antibodies directed against M. tuber-culosis as well as BCG. These antibody zones

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34 KINNMAN, LINK, AND FRYDÉN

FIG. 3. Patterns from thin-layer PAG IEF of CSF and serum from a patient with TM obtained 48 (A andB) and 65 (G through H) days after onset and the corresponding autoradiograms obtained after immunofix-ation with M. tuberculosis (C, D, I, and J) and BCG (E, F, K, and L). Arrows on A and G denote zones ofoligoclonal IgG present in CSF only, whereas the remaining arrows denote zones of intrathecally synthesizedantibodies which corresponded to some of the patient's oligoclonal CSF IgG zones found atpH values above8.6 (pH region III) and to some of the polyclonal CSF IgG zones. These zones were easily distinguished on thegel plates, but were not reproduced with desirable sharpness in the photos.

were not found in the corresponding serum spec-imens, and they were therefore considered torepresent specific oligoclonal IgG antibodies syn-thesized within the CNS. However, not all oli-goclonal IgG zones detectable by PAG IEF ofCSF from our patient with TM were identifiedas containing specific antibodies against M. tu-berculosis, and their antibody character re-mained unknown. Furthermore, intrathecallysynthesized antibodies against M. tuberculosiswere also found in polyclonal IgG zones. Theoccurrence of intrathecally synthesized specificantibodies migrating in polyclonal IgG zones hasalso been observed in, e.g., subacute sclerosingpanencephalitis (J. Shorr, B. Rostrom, and H.Link, J. Neurol. Sci., in press).

It is not known when the oligoclonal IgGstarted to appear in the CSF of our patient withTM, but the earliest available specimen, fromday 14 after onset, revealed oligoclonal IgGbands which were also found in the specimenfrom day 97, when the patient had recoveredclinically. Nor is it known how long the oligo-clonal response in CSF may persist after onsetof TM. CSF and serum obtained from a com-pletely recovered patient 1 year after the onset

of TM and studied by immunofixation with M.tuberculosis and BCG as antigens and by auto-radiography did not reveal any intrathecal an-tibody synthesis. This patient had normal find-ings on routine CSF studies on this occasion, butit is not known whether he had ever had anoligoclonal immunoglobulin response within theCNS (unpublished data).The significance of the finding of HSV anti-

bodies synthesized within the CNS and migrat-ing in polyclonal IgG zones in our patient withTM is not known. Intrathecally produced HSVantibodies migrating in polyclonal IgG zonesmay be found in patients with multiple sclerosisand also in a few normal controls (Rostrom etal., Ann. Neurol., in press). It may reflect theeffect of polyclonal B-cell activation.The diagnosis of TM is difficult and can still

only be confirmed by culture of CSF, givingresults after 4 to 6 weeks. Thus, therapy mustbe instituted on the basis of clinical findings andunspecific laboratory data. Antigen immunofix-ation and autoradiography introduced by Nordalet al. (10) have the advantage of being rapid.PAG IEF as the initial step for protein separa-tion and identification of oligoclonal immuno-

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OLIGOCLONAL IgG IN TUBERCULOUS MENINGITIS 35

globulin zones (Rostrom et al., Ann. Neurol., inpress) is superior to the agarose gel electropho-resis initially used (10), because PAG IEF hashigher sensitivity and reproducibility (Rostromet al., Ann. Neurol., in press). The possibility ofimmunofixation with M. tuberculosis or BCG,and subsequent autoradiography, as a diagnostictool should be tested on larger numbers of pa-tients with TM in order to define, inter alia, thefrequency of positive results and the earliestappearance of them. Furthermore, although theseven patients included as controls in the presentstudy were negative regarding presence of in-trathecal production of antibodies against M.tuberculosis and BCG, additional patientsshould be studied in order to establish the spec-ificity of the test.

ACKNOWLEDGMENTSThis study was supported by grants from the Swedish

Medical Research Council (project no. 3381).The technical assistance of Ingela Lilja is gratefully ac-

knowledged.

LITERATURE CITED

1. Frydén, A., and H. Link. 1979. Predominance of oligo-clonal IgG type lambda in CSF in aseptic meningitis.Arch. Neurol. (Chicago) 36:478-480.

2. Laterre, E. C. 1965. Les protéines du liquide céphalo-rachidien a l'état normal et pathologique. Editions Ar-scia, S.A., Brussels.

3. Laurenzi, M. A., and H. Link. 1978. Localization of theimmunoglobulin G, A and M, fi-trace protein and y-trace protein on isoelectric focusing of serum and cere-brospinal fluid by immunofixation. Acta Neurol. Scand.58:141-147.

4. Laurenzi, M. A., and H. Link. 1979. Characterization ofthe mobility on isoelectric focusing of individual pro-teins in CSF and serum by immunofixation. J. Neurol.

Neurosurg. Psychiatry 42:368-372.4a.Laurenzi, M. A., M. Marva, S. Kam-Hansen et al.

1980. Oligoclonal IgG and free light chains in multiplesclerosis demonstrated by thin-layer polyacrylamide gelisoelectric focusing and immunofixation. Ann. Neurol.8:241-249.

5. Link, H. 1979. Some aspects of immune reactions of thebrain, p. 12-28. In F. Clifford Rose (ed.) Clinical neu-roimmunology. Blackwell Scientific Publications, Ltd.,Oxford.

6. Link, H. 1973. Comparison of electrophoresis on agar geland agarose gel in the evaluation of gamma globulinabnormalities in cerebrospinal fluid and serum in mul-tiple sclerosis. Clin. Chim. Acta 46:383-389.

7. Link, H., and R. Muller. 1971. Immunoglobulins in mul-tiple sclerosis and infections of the nervous system.Arch. Neurol. (Chicago) 25:326-344.

8. Link, H., M. Panelius, and A. Salmi. 1973. Immuno-globulins and measles antibodies in subacute sclerosingpanencephalitis. Arch. Neurol. (Chicago) 28:23-30.

9. Link, H., and G. Tibbling. 1977. Principles of albuminand IgG analyses in neurological disorders. III. Evalu-ation of IgG synthesis within the central nervous systemin multiple sclerosis. Scand. J. Clin. Lab. Invest. 37:397-401.

10. Nordal, H. J., B. Vandvik, and E. Norrby. 1978. Dem-onstration of electrophoretically restricted virus-spe-cific antibodies in serum and cerebrospinal fluid byimprint electroimmunofixation. Scand. J. Immunol. 7:381-388.

11. Tibbling, G., H. Link, and S. Ohman. 1977. Principlesof albumin and IgG analyses in neurological disorders.I. Establishment of reference values. Scand. J. Clin.Lab. Invest. 37:385-390.

12. Tourtellotte, W. W. 1975. What is multiple sclerosis?Laboratory criteria for diagnosis, p. 9-26. In A. N.Davison et al. (ed.), Multiple sclerosis research. Elsev-ier-North Holland Publishing Co., New York.

13. Vandvik, B., and E. Norrby. 1973. Oligoclonal IgGantibody response in the central nervous system todifferent measles virus antigens in subacute sclerosingpanencephalitis. Proc. Natl. Acad. Sci. U.S.A. 70:1060-1063.

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