Craig Douglas and Dylan Rigby, who are both from Essex, are charged with promoting a lottery and advertising unlawful gambling.
Mr Douglas makes gaming videos on YouTube under the pseudonym Nepenthez.
He is also charged with inviting children to gamble.
The two men appeared at Birmingham Magistrates' Court. The case has been adjourned until 14 October.
The Gambling Commission, which brought the prosecution, has been looking into the rise of video game gambling.
It is warning parents that children can be drawn into betting with so-called skins - virtual goods such as weapons or clothes that are a feature of many popular games.
It has been estimated that the global market in betting on video games is worth as much as Â£4bn.

The New Syrian Army (NSA) said it had captured a number of IS positions on the outskirts of Albu Kamal, after overrunning a nearby military airport.
However, a raid into the town launched at dawn is reported to have been repelled by militants.
The offensive, which began on Tuesday, is aimed at cutting a key route between IS-controlled areas in Syria and Iraq.
The NSA said it was co-ordinating the assault with Iraqi government forces, who were advancing on the border from the other side.
Several hundred rebels are reportedly involved in the attack on Albu Kamal, just a few kilometres from the Iraqi border in Deir al-Zour province.
The NSA and the Syrian Observatory for Human Rights, a UK-based monitoring group, both said on Wednesday morning that the rebels had taken control of Hamdan airport, about 5km (3 miles) north-west of the town.
Fighting was continuing between the two sides in agricultural areas around the town, and coalition warplanes were conducting air strikes on IS hideouts in the town, according to the Syrian Observatory.
NSA spokesman Mozahem al-Saloum told the Associated Press that airborne fighters had also been dropped onto the southern edge of Albu Kamal.
Later on Wednesday, the NSA issued a statement saying its forces had "conducted a deep penetration raid" into Albu Kamal.
"At this time fighting is still ongoing, but the NSA maintains control of the desert, the approaches to Albu Kamal, and maintains freedom of manoeuvre," it added.
The Reuters news agency cited two rebel sources as saying that the NSA had been forced to retreat to the outskirts after an IS counter-attack.
One of the sources said militants had encircled the rebels in a surprise ambush. The rebels had incurred heavy casualties and lost some of their weapons, the source added.
As the rebels advanced on Albu Kamal, IS released a video showing militants beheading five young men in the town who they claimed were working with the NSA, the Syrian Observatory said.
The NSA was formed about 18 months ago by rebel factions driven out of eastern Syria by IS, which proclaimed the creation of a "caliphate" in June 2014.
NSA fighters, who have been trained at US-run camps in Jordan, captured the Tanf border crossing between Syria and Iraq from IS earlier this year.
IS is also facing pressure from a US-backed offensive in northern Syria by an alliance of Kurdish and Arab fighters, who have surrounded the town of Manbij and cut a route to the Turkish border from the IS stronghold of Raqqa.
In a separate development on Wednesday, at least seven people were killed in a car bomb attack in the Kurdish-controlled northern town of Tal Abyad, near the Turkish border, state media reported.
Two men have appeared in court charged with offences under the Gambling Act in what is believed to be the first prosecution involving betting on video games.
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US-backed Syrian rebels say they are closing in on a town on the border with Iraq held by so-called Islamic State.