Jpn. J. Infect. Dis., 53, 1561161, 2000

Original Article

Jackfruit Lectin: Properties of MitogenlClty and the Inhibitionof Herpesvirus Infection
Nuanchawee Wetprasit* , Weena Threesangsri 1 , Natapong Klamklail
and MontriChulavatnatol2
Department ofBiotechnology, FacultyofScience, Ramkhamhaeng UniversiO!,
Bangkapi, Bangkok I 0240,

JNational Institute of Health, Department of Medical Sciences, Ministyy of Public Health.
Nonthaburi 11000 and

2Department of Biochemistry, Faculo, of Science, MahidoI UniversiO/,
Rama 6 Road, Bangkok 10400, Thailand(Received March 30, 2000. Accepted August 2 1 , 2000)
SUMMARY: Jack fruit lectin (JFL)from Arloca.rpus hete.nophyllus has been found to exhibit inhibitory activity invitrowith a?ytopathic effect towards herpes slmPlex vlruS type 2 (HSV‑2), varicella‑zoster yiruS (VZV), andcytomegalovlruS (CMV). The 50% inhibitory dose values from plaque reduction assay (inactlVation) were 2･5,
5, and 10 LLg/ml of JFL for HSV‑2, VZV, and CMV, respectively. Lymphocyte proliferation was slgnificantly
increased in the presence of the JFL in the concentration range of2.5 to 50 LLg/ml, but was reduced at 500 LLg/

ml. It was found that CD16'/CD56'cells (natural killer cells) were induced among the primary lymphocyte

subpopulations. The activityofnatural killer (NK) cells was not affected by JFL in the concentration range of 5
to 500 LLg/ml. These data suggest that JFL is mitogenic for NK lymphocyte (CD16'/CD56') and also active

agalnSt HSV‑2, VZV, and CMVa tetramer of Mr 62,000 and consists of two subunitswith Mr

INTRODUCTION

of 1 8,000and two subunitswith肋of 13,000, whereasJaCalin

Lectins are carbohydrate‑binding proteins of non‑immune

is an a282 tetramer of Mr 39,000‑54,000 (14,17‑19). JFL

origin that are o洗en derived from plant seeds. They are well

can agglutinate human, rat, mice, sheep, and goat erythrocytes

known to have biologlCal activity ln relation to lymphocyte

as well as human and rat spem (16,20). Fu仙emore, it has

proliferation (1 ,2), the inhibition of virus infection (3,4), andalso been fわund to agglutinate several glycoproteins inbacterial agglutination (5,6). The binding of lectins to cell or

biological nuidsand canbe used to agglutinate several serovarsof Bacillus thuringiensis, serovar H4 (sotto), and Salmonella

viral glycoproteins and glycolipids depends on their sugar

specificities (7)･ Lectins have previPusly been demonstrated
to reduceviral infectivlty ln eXPerlmentS uSlng Cell mon0‑
1ayers･ For.example, concanavalin A (Con A) and wheat gem

JaCalin, it does differ in some of its biological properties. To

agglutinatlnin (WGA) inactivate and reduce rubella virus

further establish the biologlCal properties ofJFL, the in vitro

multiplication (3 ,8). Ⅵmses that contain glycoproteins in theirinhibitory effects of JFL towards HSV12, varicella‑zoster

りphi, except for S. pwaりphi, Escherichia coli, B. Sphaericus,

and Sacchay10myCeS CereVisiae. Although JFL is similar toenvelopes would be expected to interactwith lectins sincevirus (VZV), and cytomegalovirus (CMV) were studied.
Moreover, its actions on lymphocyte proliferation and naturalkiller (NK) activity were also investigated.

lectins have a specific affinity for their carbohydrates. The
inhibition of virus infection after treatment with lectins of

Con A (9,10), Limulus polyphemus (LPA) (8), WGA (8),Bauhinia purpurea (BPA), and Eranthis hyemalis agglutinins

MATERIALS AND METHODS(1 1) has been reported for herpes.simplex virus (HSV)･ In
addition, the replication of rabies vlruS is successfully inhib‑

Cells: BHK‑21 (baby hamster kidney) and MRC‑5 (humanited by WGA and LPA (12).
Mitogenic stimulation is an effect caused by the interac‑

embryonic lung) cells were cultured at 37oCinEagle's minimaltion or lectins with cells. It plays a role in slgnal transduction

containing 1.2 g/1 NaHCO3 and supplemented with 1 00 U/ml

at the surface of lymphocytes for growth and proliferation.
Many lectins have been recognized to be mitogenic. Jacalin

penicillin, 50 jig/ml streptomycin, and 10% or 4% (vol/vol)fetal bovine sera (FBS) (Biowhittaker, Walkersville, Md. , USA)

from Artocarpus integrlfolia is alsomitogenic to CD4'T cells

for growth medium and maintenance medium, respectively.

(13,14) and blocks in vitro human immunodeficiency virus

type 1 (HIV‑1) infection to a T‑lymphocyte celHine (15).

Virus: The HSV‑2 (LB‑strain) and CMV (AD‑ I 69 strain)
were propagated in BHK‑2 1 and MRC15 cells, respectively.

Recently, jack fruit lectin (JFL), derived from the seeds ofA.

Virus stocks were then obtained by freezlng and thawlng at

essential medium (MEM) (Nissui Phamaceutical C0. , Tbkyo)

heterophyllus, which is a tropical tree of the family Moraceae‑70oC and 370C to break the cells, followed by centrifuga‑

found in Thailand, has been found to be similar to Jacalin intion at l,200 g fわr 1 0 min. The supematant was collected as

being specific for N‑acetylgalactosamine (16). This lectin isvirus stock seeds and kept frozen at ‑700C until use.

The VZV (Kawaguchi strain) was adsorbed to MRC‑5 cells*Corresponding author: Tel/Fax: +66‑2‑319‑4385, E‑mail:in a 751cm2 tissue‑culture mask at 37oC for 60 min. The main‑

nuanwee@yahoo. com

tenance medium was added, and the culture was incubated156

until an 80% cytopathic effect (CPE) was observed. Each

Preparation of peripheral blood plOnOnuCIear cells

infected culture was mixed with an uninfected one in a ratio(PBMC): Mononuclear cells were separated from theof I :4 (vol/vol) according to a trypsin.ization method toheparinized blood by denslty gradient centrifugation uslng

prepare cell‑freeviruS, as described prev10uSly (2 1 )･

Ficoll‑Hypaque (Isoprep, Robbins Scientific Corp. , Surmyvale,

Purification of JFL: JFL was purified from the seeds ofCalif･, USA) (22)･ The collected mononuclear cells w.ere

A. heterophyllus according to a previous study (16)･ Seeds

washed three times with RPMl 1640 (Flow LaboratorleS,

were chopped and homogenized fb∫ 3 min in 3 volumes of 5

McLean, Va., USA) by centrifugation at 400 × g for 10 min

mM phosphate‑buffered saline (PBS), pH 7･4, using a Waring

at 4oC. The mononuclear cells were counted and adjusted to

Blender. The following Steps Were Performed at 4oC･ Afteran appropriate concentration in complete RPMI 1640 (RPMI
1640 medium supplemented with 2 mM glutamine, 10 mM

the homogenization, the mixture was sti汀ed fわr 2 h and then

HEPES, 100 U/ml penicillin G, and 100 jig/ml streptomycin
containing lO% FIiS)･
Lymphocyte proliferation response assay: 100 Ill ofcentrifuged at 3,000 X g for 15 min to remove insoluble debriS･
The supematant was then collected, and ammonium sulfate

was added to 60% saturation. The precIPltate Was COllectedby centrifugation at 27,000 X g for 15 min, resuspended in

PBS, and dialyzed for 24 h with repeated changes ofbuffer･

prepared mononuclear cells (2 X 106 cells/ml) we,e cultured
ln triplicate in 96‑well microtiter plates (Costar, Cambridge,

The dialyzed fraction †as applied to a column (1 ･8 × 6･5 cm)

Mass., USA)with 100 LL1 0fcomplete RPMI 1640 containingofルーacetylgalactosamlne‑agarOSe (Sigma Chemical Co･, St･different concentrations of JFL or phytohemagglutinin (PHA)0.03 M galactose in PBS, and the absorbance of the collected

HA 16/17 (Murex Diagnostics Ltd., Dartfbrd, England) at 1
jig/ml (final concentration). The cultures were incubated at
37oC with 5% CO2 for 72 h. Eighteen hours before harvest,

仔actions was monitored at 280 nm. The galactose‑eluted丘.ac‑

20 LEI of 0.5 JLCi 3H‑thymidine (specific activity, 8･3 mCi/Louis, Mo., USA) that was pre‑equilibrated with PBS. Afterwashing off the unbound material, the column was elutedwith

tions exhibiting hemagglutination activity(using 2 lvol/vol]plg‥ Amersham, Buckinghamshire, UK) was added･ After

sheep erythrocyte suspension) Were pooled and concentrated

Incubation, cells were washed with a multichamel automaticfor use as JFL in this study.

cell harvester (Dynatech Lab., Inc., Sussex, UK) onto glass‑

AntiviraJ activity assay: JFL was studied for its antiviralfiber filters (Whatman International Ltd., Maidstone,

activity by fbllowlng plaque reduction under three conditions･England). The 3H‑thymidine incorporation was measured by(1) Pre‑treatment condition: BHK‑21 and MRC‑5 cells

a liquid scintillation counter (Wallac Oy, Turka, Finland). The

were pre‑incubated at 37oC for 2 h with various concentra‑

degree ofactivation was expressed as a stimulation index (S･I･),

tions of JFL in the maintenance medium. A鮎r the pre‑i.ら., the ratio of 3H‑thymidine uptake in counts per minutetreatment, the monolayer cells were washed two times with(cpm) of samples with to those without lectin･
Flow cytometry analysis: The prepared mononuclear cells

the medium and then inoculated at 37oC for 60 min with 100

plaque‑forming units (PFU) of virus. Any unadsorbedvirus(I X 107 cells/ml) were culturedwith JFL or PHA in complete

was discarded, and the cells were washed with the medium.

RPMI 1640 Containing 10% FBS. A鮎r a 24‑h incubation,

The monolayers were仇en incubated in the medium until 80%

cell pellets were haⅣested and washed with RPMI 1640 by

CPE was observed in the control containlng nO JFLI The HSV‑

centr血gation at 400 × g f♭r 5 min at room temperature･ Cells

2 CPE was measured by the neutral red assay, whereas thatwere stained uslng fluorescein‑conjugated CD3 , CD3/CD4,
CD3/CD8, CD3/CD1 6+56, or CD3/CD19 mouse monoclonal
antibodies (Becton Dickinson Immunocytometry System, San
Jose, Calif., USA). After staining, the cells were fixed in PBS
containlng 3% formaldehyde, and the fluorescence intenslty
was measured by now cytometry using a FACScan (Becton
Dickinson).

ofVZV or CMV was measured by stainlng With methylene
blue.

(2) Post‑treatment condition: BHKl2l or MRC‑5 cells
were adsorbedwith loo PFU of virus at 37oC for 60 min; theunadsorbed vims was then discarded, and the cells were
washed with仇e medium. After washing, the monolayer cells

medium. The infected cell cultures were incubated until an

Assay ofNK activity: The PBMC (2 X 106 cells/ml) Werewashed and resuspended in complete RPMI 1640 and incu‑

80% CPE was obseⅣed in the control without any treatment.

bated with or without JFL or PHA at 37oC with 5% CO2 fわrwere incubated with various concentrations of JFL in the

18 ‑ 24 h. K562 cells, kindly provided by Dr. Busarawan
Srivanna (National institute of Health, Department of Medical(3) Inactivation condition: 100 PFU of viruS.was mixed
with the medium contaimng various concentrat10n岳of JFL

and incubated at 37oC for 30 min. After incubation, 0.2 ml

Sciences, Thailand), Were used as target cells and were grown

was inoculated into monolayer cells. The inoculum wasin complete RPMI 1 640 Containing 1 0% FBS･ The target cells

allowed to be adsorbed to monolayer cells fわr 60 min at 37oC･(1.5 X 106 cells) were labelled with Na251CrO. (specific

The unadsorbed vims was then discarded, and the cells wereactivity, 37.0 MBq/mg, Amersham) at 37oC, 5% CO2 fわr 45

washed with the medium. The cells were incubated with themin and washed three times with RPMI 1640 Containlng 10%

mediumunti1 80% CPE was observedinthe control cell culture

FBS.

The cytotoxicityassay was performedinU‑shaped microtiter(Without JFL).

In order to examine the effects of sugars on the JFL inhibi‑tion of virus infectivity, the galactose, N‑acetylgalactosamine

plates (Corning lnc., Corning, N.Y., USA) using 1 X 104
target cells/well and PBMC in varylng ratios･ After adding

or glucose (Sigma), was mixed with JFL (40 LLg/ml final

the target cells, the mixture was incubated for 4 h at 37oC

concentration) and each virus (100 PFU). After incubation

with 5% CO2. The plates were then centrifuged at 200 X g

for 30 min at 37oC, themixture was inoculated into a monolfor 5 min. The supematant fraction from each well (100上土1)

Blood samples: Blood samples were collectedfrom healthy

was transferred into a tube, and the radioactivitywas counted
in a Gamma counter (Packard lnstrumental Co., Meriden,

Thai adults who had no history of hepatitis B infection orConn･, USA)･ The percentage of cytolysis †as calculated

layer cell culture and processed as above･

according to the fbmula: %cytolysis = (expenmental release

exposure to HIV1 1 , antimalarials, corticosteroids, or immuno‑‑ spontaneous release) / (maximal release ‑ spontaneous

suppressive dmgs･157

release). Spontaneous release was measured after incubationor the target cells with the medium alone, while maximal

jig/ml ofJFL (P< 0.05). However, at 500 LLg/ml ofJFL, an
inhibition was observed. In contrast, PHA at 1 LLg/ml stimu‑

release was measured after lysIS Ofthe target cells with 5%

lated PBMC proliferation by approximately 300‑fold. In

Triton x‑loo. NK cell activlty Was expressed as lytlC units

order to define the subpopulation among the lymphocytes(LU)/107 PBMC as detemined by least squares analyses ofthe percentages at different cell ratios. One LU was definedas the number ofNK cells required for 20% specific lysIS Of

rl山■

0 80 餌 仰 20 0(%)LJO慧9!)d!tlnNSn.JI^

1 X 104 target cells･
Statistical analysis: Data were expressed as the mean ±the correspondent standard deviation (SD), and statisticalanalysts Was Carried out by employlng the Student's t‑test.

RESUIXS

Antiviral activity of JFL: To determine the antiviral
activityof JFL, three experimental conditions (pre‑treatment,

0

10

0

1

0

20

0

10

20

20

班

仰

post‑treatment, and inactlVation) were used in the plaque‑reduction assay. JFL was found to inhibit the CMV‑induced
LIL

plaque reduction under all three conditions. The plaque(%)uo!le3([d!tJnHSN!^∞ 紛 餌 仰 20 0

reduction by HSV12 was not affected by pre‑treatment with
JFL, and the plaque fbmation induced by VZV was only

affected in the inactivation assaywith the 50% inhibitory dose
ofJFL 5 LLg/ml (Table 1 and Figure). The inhibitory effectofJFL on the infectivityofthe virus was diminished by theaddition of 0.2 M galactose or N‑acetylgalactosamine but not

glucose. Each sugarwithout JFL showed no effect on thevirus‑induced plaque formation (Table 2).

Lymphocyte proliferation response and subpopula‑30

40

tioms: The mitogenic potential or JFL was investlgated byuslng PBMC. PBMC were exposed to different concentra‑Ll(%)uo葛3!ldZlLnHSnJ!^

tions ofJFL before the proliferative response was measured

08642 000000

by a DNA synthesis assay. The results are shown as S.Ⅰ. in

Table 3. It was found that JFL induced proliferative responses.

The induction was small in the presence ofO.5 ng/ml to 0.5
LLg/ml ofJFL, but was significant in the range of2.5 to 50

Table 1. lnhibitory dose ofJFL in plaque formation in thepresence of a virus
Treatment

HSV‑2
Pre

̲trea

30

40

50% inhibitory dose of JFL ( 〝g/ml)1

tment

NA2

Post‑treatment

NA
3

Inactivation

VZV

5

2. 5

N

JFL Concentration (LAg/mI)

CMV‑ tJ 1 5 5 0

Figure. Dose‑response curves ofHSV‑2 (A), VZV (B), and CMV (C)multiplications in various concentrations of JFL by plaque reduction

A

assay after pre‑treatment ( ◆ ), post‑treatment ( ● ), and inactivation

5

1Minimal non‑cytotoxic concentration ofJFL = 125 JLg/m1(▲ ) assay.

2NA: No activity〝=3

Table 3. Effect ofJFL on the proliferation of peripheral bloodmononuclear cells (PBMC)
Table 2. Inhibitory effect ofJFL on the infectivity (plaque formation)JFL

orHSV‑2, VZV and CMV by inactivation assay
HSV‑2

VZV

Final concentration
orJFL (〝g/ml)

No. of No. of

No. orplaques plaquesplaques150

3H‑thymidine

Stimulation

index(S.I.)1

incorporation (cpm) I

None1245±1841

O.5 ng/m1

1415±1921.21±0.16

5 mg/m1

1659±1801.43±0.50

100

50 mg/m1

1710±160

1.46±0.57

0750700.5 〝g/m1

1853±136

1.57±0.66

2.5 ug/m1

3398±2572

3.23±1.372

hXU Oノ 0ノ ーー

40+GalNAcl

148

40+Glut
0+Gal

4387±2932

4.66̲+2312

67249± 33932

72.99±39.072

500 LLg/ml

0+GalNAc

0+Glu

5 〟g/m1

50 LLg/m1

PHA (1 LLg/ml)
145

105

291 ±642

0.33±0.182

238227± 138632

300.5±207.892

lEach value represents mean ± SD

1Final concentration = 0.2 M2J}< 0.05

fl=3

〝=10

158

Table 4. Effect ofJFL on thepercentage ofCD3+, CD4', CD8十, CD16十/CD56'and CD19'

subpopulation cells in nomal human donors
JFL CD3+

CD4+

CD8+

CD 1 6+/CD56+59.4± 10.441

3 1.6±4.92

24±6.78

0.5 jig/m1

57.2±9.33

30.2±4.87

23.2±3.66

31 ±9.08

13.8±6.43

5 LLg/m1

53±6.8426.2±4.26

21.2±2.71

32.6±10.212

18.8±6.62

50 LLg/m1

51.8±5.84

22.6±5.92

21±7.0134.6±9.832

14.8±5.31

None26.2±10.19

CD 1 9十12.6±5.75

tEach value represents mean ± SD2f〉< 0.05〃=5

Table 5. Effect of PHA on thepercentage ofcirculatlng CD3', CD4', CD8+, CD1 6'/CD56'
and CD1 9+ subpopulation cells in l10mal human donors
PHA CD3+

CD4+

None

60.8±3. 191

CD 1 6+/CD56+37.8±6.72

23.2±4.35

58±4.0538±6.2322±4.5657.8±2.23

37.2±4.71

22.6±3.98

0.2 LLg/m1

1 FLg/m1CD8十23.8±1.94

CD 1 9+11.4±1.74

29.8±1.472

11±2.1031.4±3.012

11.2±2.56'Each value represents mean ± SD

2J) < 0.05〝=5

Table 6. Activityofcells with natural killer (NK)function in

None

90.56±22.941

5 Fig/m1

74.33±8.12

been found to be effective agalnSt Viral replication after
treatment (8,25,26). The antiviral activity of JFLinresponse
to HSV‑2 and CMV either before or after viral infection ofcell monolayers was observed at different doses (Table 1);
this result differs markedly from the lack of effect reported

50 〟･g/m1

85.71±8.82

for collectin, mannan‑binding protein, and bovine conglutinin

500 〟g/ml

84.09± 12.51

on HSV‑2 (27). These results suggest that JfL may act either

PHA 0.2 〃･g/ml

102.59±30.92

lytic units (LU)/I 07 PBMC in the presence ofJFL extract

Lectin concentrations LU/I 07 PBMC

JFL

l ′上g/mlon the surface of host cells or directly on theviral envelope,

thereby inhibiting viral infectivity.
This is the first report of the effects ofJFL on lymphocyte

277.13±81.452

lEach value represents mean ± SD

2p< 0.05

proliferation. The degree of lymphocyte reactivity, S.I. , which〝=5

wFs higher than the control (Table 3), su.ggests that JFL canstlmulate the PBMC, i.ら., that JFL is a mltOgen. However, athigh concentrations JFL was found to decrease lymphocyte
stimulated by JFL, an analysis of lymphocyte subsets uslng aproliferation responsiveness , suggestlng that JFL preparations

now cytometer was carried out. A major Significant increase

are tofic to cells at high concentrations (500 LLg/ml) due toin the percentage orCD16+/CD56+ was obseⅣed at 5 and 50

the stlmulation of PBMC. Lymphocyte proliferation underlectin stimulation has been fわund to produce optimal levels.LLg/ml ofJFL (P < 0･05) (Table 4), and similar significant

Increase Was induced by themitogenic PHA (Table 5).

or mitogenic activity (28). According to our study of JFL

NK activity: NK activity in the presence ofJFL was

stimulated lymphocyte subsets, there appears to be a change

assessed and compared with that in the presence of PHA. A
slgnificant induction of NK activlty aS measured in LU waswhich were NK cells (Table 4). This finding differs from thatin the numbers or CD 1 6+/CD56+ lymphocyte subpopulations

demonstrated through the use of PHA‑treated PBMC. In
contrast, no increase in NK activity was detected using JFL
treated PBMC (Table 6).

of in vitro studies showlng thatjacalin stimulates the CD4十lymphocyte (14,28) and B‑cell prohferation at the highconcentration of 2 mg/ml (28). Other lectins purified fromseeds of different Artocarpus species in Vietnam (A.
asperulus, A. hetey10Phyllus, A. musticata, A. me/inoサIus, A.

DISCUSSION

parva, and A. petelotii) have also been reported to stimulate
Several p)ant lectins have been shown to inhibit the infec‑

human PBMC proliferation with variable mitogenic efficien‑

tivityofviruses. For example, jacalinhas been found to inhibit
in vitroinfection of HIVl 1 without preventlng the virus from

Furthemore, Con A, poke weed mitogen (PWM), and PHA

binding to the host cell (15,23). In this study, the in vitro

have been fわund to be mitogens that can induce Ii cells toantiviral activity of JFL was demonstrated for the first time.

proliferate in the presence of 1 to 5% ofT cells (30). These

Previously, many lectins with different specificities have beenlectins activate B cells to proliferate indirectly, as theycies, but they all stimulate only the CD4+ T lymphocyte (29).

proven to inhibit viral infection. In the case of PHA, theinduce T cells to produce mitogenic factors actlng On B cells

treatment of CMV pr10r tO the infection of cell monolayers
reduced theviralinfectivity by 90% and a 99% reduction in(31,32). In the present study, PHA was fわund to induce a

infectivity was obtained by pretreatment of cellswith PHA

did not alter the NK cell activity (Table 6). The observed

slgnificant increase in the function ofNK eel)S, whereas JFL(24). The inactivation ofHSV‑ 1 adsorption has been observed

inhibitory effect of PHA on the cytotoxicityof NK cells may

in respohse to Con A, succlnyl‑Con A, WGA, and helix

be caused by the production of some cytokine such as

pomatia agglutimin, whereas BPA and soybean agglutinin haveinterleukin‑4 (IL‑4) (33). However, JFL is different from PHA

159

selective inhibitors of human immunodeficiencyvirus
and cytomegalovirus replication in vitro. Antiviral Res.,in this respect.

The present study has demonstrated that JFL is a new18, 191‑207.

mitogen仇at is mitogenic toward CD 1 6十/CD56十cells･ Hence,

JFL can be used instead of other well‑known mitogens such

12. Conti, C., Superti, F. and ¶iiang, H. (1986): Membraneas Con A, PHA, and PWM to specifically activate lymphocyte

carbohydrate requlrement fわr rabies virus binding toproliferation. However, the results obtained h this study

chicken embryoィelated cells. InteⅣirology, 26, 1 64‑ I 68.

preliminarily show JFL to have the characteristic or in vitro

1 3. Pineau, N.,Acouturier, P., Brugier, J. C. and Preud'horrune,

immunomodulating activlty･ Further studies should be

J.し. (1990): Jacalin: a lectin mitogenic fわr human CD4

T lymphocytes. Clin. Exp. lmmuno1., 80, 420‑425.

perfわrmed to determine whether JFL exhibits any other

14. Aucouturier, P., Pineau, N., Brugler, J. C., Michaesco,

immunostimulatory activlty.

E., Duarte, F., Skvaril, 已 and Preud'homme, J.し. (1989):

Jacalin: a new laboratory tool in immunochemistry and

ACKNOWLEDGMENTS

cellular immunology. ∫. Clin. Lab. Anal., 3, 244‑251.

We are especially grateful to Dr. Busarawan Sriwanthanafor her support, excellent technical assistance, and helpful

1 5. Favero, J., Corbeau, P., Nicolas, M., Benkirani, M., Trave,G., Dixon, J. F., Aucouturier, P., Rasheed, S., Parker, J.

discussions. We would also like to thank Dr.Watana Auwanit

W. and Liautard, ∫. P. (1993): Inhibition or humanimmunodeficiency virus infection by the lectin JaCalinfor his help and support.

and by a derived peptide showlng a Sequence Similarlty
with gp120. Eur. ∫. Immuno1., 23, 179‑185.

REFERENCES

16. Na叩Juntra, P., Muanwongyathi, P. and Chulavatnatol,

1. Lis, li. and Sharon, N. (1986): Biological properties ofM. (1985): A sperm‑agglutinating lectin from seeds oflectins. p. 270‑277. In LE. Liener, N. Sharon and I.J.
Goldstein (eds.), The Lectins: Properties, Functions andjack fruit (Artocarpus hetey10Phyllus). Biochem. Biophys.

Res. Commun., 128, 833‑839.
171 AppukuttFn, P･ S･ and Basu, D･ (1985): Four identical

Applications in Biology and Medicine. Academic Press,

subunits ln jack fruit seed agglutinin offer only two

Orlando, Fla., USA.

saccharide binding sites. FEBS Lett., 180, 33 1‑334.

2. Stobo, ∫. D., Rosenthal, A. S. and Paul, W. E. (1972):

18. Aucouturier, P., Mihaesco, E., Mihaesco, C. andFunctional heterogenelty Of murine lymphoid cells. Ⅰ.

Responsiveness to and surface binding of concanavalin

Preud'homme, ∫.し. ( 1 987): Characterization ofjacalin,

A and phytohemagglutinin. ∫. Immuno1., 108, 1‑17.

the human IgA and IgD binding lectin from jack fruit.

Mol. lmmuno1., 24, 503‑511.

3. Urade, M., Sato, M., Yoshida, H., Shirasuna, K., Miyazaki,
T･ and YamaPoto, N･ (1978)‥ Effect of con?anavalin A

19. Ahmed, H. and Chatteljee, B. P. (1989): Further character‑on the infectlVlty Ofrubellavirus and its varlantS. Arch.
Viro1., 56, 359‑363.

ization and immunochemical sbdies on仙e carbohydrate

specificity ofjackfruit (Artocarpus integrlfolia) lectin.

4. Weiler, B. E., Schroder, H. C., Stefanovich, V., Stewart,D., Forrest, J. M. S.,Allen, L. B., Bowden, B. J., Kreuter,

20. Prapunpoj, P. and Chulavatnatol, M. (1996): Interaction

M･ H･, Moth, R･and Muller, W･ E･ G･ (1990)‥ Sulphoevemap,

between jack fruit lectin and rat epididymal sperm. J. Sci.

J. 丘iol. Chem., 264, 9365‑9372.

Soc. Thailand, 22, 21‑30.

a polyanionic polysaccharide, and the narcissus lectln

potently inhibit human immunodeficiency virusinfec‑

21. Yamanishi, K., Matsunaga, Y., Ogino, T., Takahashi,tion by binding to viral envelope protein. ∫. Gen. Viro1.,

M･ and TakamizaTa, A･ (1980)‥ Virus replication and71, 1957‑1963.

localization of varlCella‑zoster vims antlgenS in humanembryonic fibroblast cells infectedwithcell‑free virus.

5. Cole, H. B., Ezzell, ∫. W, Keller, K. F. andDoyle, R. ∫.

Infect. Immun., 28, 536‑541.

(1 984): Differentiation of Bacillus anthracis and other

Bacillus species by lectins. J. Clin. Microbio1., 19, 481

22. Boyum, A. (1966): Separation ofleukocytes from blood53.

and bone ma汀OW. Scan. ∫. Clin. Lab. Invest., 21, Suppl.

97.

6. Davidson, S. K., Keller, K. F. andDoyle, R. ∫. (1982):

23. Corbeau, P., Pasquali, J. L andDevaux, C. (1995): Jacalin,

Differentiation of coagulase‑positive and coagulase‑
negative staphylococci by lectins and plant agglutina‑

a lectin interacting with 0‑linked sugars and mediatingtions. ∫. Clin. Microbio1., 15, 547‑553.

protection of CD4+ cells agalnSt HIV‑1, binds to the

7. Lis, H. and Sharon, N. (1986): Lectins as molecules andas tools. Annu. Rev. Biochem., 55, 35‑67.

extemal envelope glycoprotein gp120. Immunol. Lett.,
47, 141‑143.

8. Marchetti, M., Mastromarino, P., Rieti, S., Segani, L. and24. Ito, M., GiⅣin, L and Ba灯on,A.し. (1978): Inactivationof human cytomegalovirus by hemagglutinin. Arch.
Viro1., 57, 97‑105.

Orsi, N. (1995): Inhibition of herpes simplex, rabies andrubellaviruses by lectinswith different specificities. Res.
Viro1., 146, 21ト215.

25. Hansen, ∫. E., Nielsen, C. and Vestergaard, B. F. (1991):

10. Ito, M. and Ba灯on, A.し. (1974): Inactivation ofhe叩eS

Inl1ibition of human.immunodeficiency yirus 1 (HIVl 1 )and herpes sipIPlex vlruS (HSV‑ 1 ) infectivltywith a broad
range oflectlnS. Scand. J. Infect. Dis., 23, 425‑430.
26. Garrity, P., Szelc, C., Paquette, C., McEvoy, M., Millette,

simplex vims by concanavalinA. ∫. Viro1., 13, 13 12‑13 18.

R. and Adler, R. (1982): Inhibition of he叩eS Simplex

11･ Balzarini, J., Neyts, ∫., Schols, D., Hosoya, M., Van

Damme, E., Peumans, W. and De Clercq, E. (1992): The

vims replication by succlnyl concanavalin A. Antimicrob.
Agents Chemother., 21 , 450‑455.

mannose specific plant lectinsfrom Cymbidium hybrid

27. Fischer, P. B., Ellermann‑Eriksen, S., Thiel, S., Jensenius,and Eplj,actis helleborine and the (N‑acetylglucosamine)n‑Specific plant lectin from Urtica dioica are potent andprotein and bovine conglutinin mediate enhancement of

9. Oka血, Y and Rim, ∫. (1972): hcubation orconcanavalin

A with enveloped viruses and host cells. Virology, 50,
507‑515.

J. C. and Mogensen, S. C. (1994): Mannan‑binding160

herpes simplex virus type 2 infection inmice･ Scand･ J.

subpopulatios. I. quantitative isolation of human T and

Immuno1., 39, 439‑445.

B cells and response to mitogens. Hmmunol･, 1 13, 1 1 13‑

28. Bunn‑Moreno, M. M. and Compos‑Neto, A. (1981):

1121.

Lectin(S) extracted from seeds ofArtocarpus integrlfolia

31. Brochier, J., Samurat, C., Gueho, J. P. and Revillard, J.P.

Oackfruit): plant and selective stimulator(S) of distinct(1976): T‑dependency of human B lymphocyte prohf‑

human T and B cell functions. J. Immuno1., 127, 427‑

erative response to mitogens. Int. Arch. Allergy Appl･

429.

Immuno1., 51, 10ト116.

32. Stashenko, P. P. and Schlossman, S. F. (1977): Antigen

29. Blasco, E., Ngoc,し. D., Acouturier, P., Freud Homme,

J. L and Barra, A. (1996): Mitogenic activity of newrecognlt10n: the specificity of an isolated T lymphocyte

lectins from seeds of wild Artocarpus species from

population. ∫. Immuno1., 1 18, 544‑550.

Vietnam. Comptes Rendus Acad. Sci･ Ill, 3 19, 405‑409･

33. Banchereau, ∫. and Rybak, M. E. (1994): Interleukin‑4.

30. Chess, L., MacDermott, R. P. and Schlossman, S. F. (1974):

p. 99‑126. In A.W. Thomson (ed.), The Cytokine Hand‑

book. Academic Press Inc., San Diego, Calif., USA.

Immunologic functions of isolated human lymphocyte

161