Apart from Khosla, those who made the cut include outsourcing firm Syntel's founder Bharat Desai, venture capitalist Kavitark Ram Shriram and software king Romesh Wadhwani.
While Khosla occupies the 308th spot in the list with fortunes worth $1.3 billion, Desai ranks 252 with a net worth of $1.6 billion.
Shriram occupies the 288th position with assets worth $1.45 billion and Wadhwani is on the 290th spot in this year's list with a total valuation of $1.4 billion.
The list is topped by Gates, which has retained his position as the richest man in America with a net worth of $54 billion, for the 17th year in a row.
Desai earned his engineering degree from IIT Mumbai and moved to the US after landing a programming job for Tata Consultancy Services in 1976.
He founded outsourcing outfit Syntel in 1980 with wife, Neerja Sethi, while earning his MBA from University of Michigan. Although he stepped down as Chief Executive last year, Desai remains the Chairman.
The Florida resident launched a deep value hedge fund last year and recently established foundation supporting entrepreneurship and education.
Desai played for India in 1994 bridge world championship and says "his biggest failure" is "not being able to play cricket for India."
Shriram is the founding board member of Google and one of the first investors in the company. "Shriram was one of the first people to write a cheque to Google co-founders Sergey Brin and Larry Page in 1998.
"He would advise them one day a week in their Menlo Park garage," Forbes said.
The University of Madra alumnus is still a board member and large shareholder of the search giant. The California-resident is now running his own investment firm, Sherpalo Ventures, which has big stakes in online outsourcer 24/7 Customer.
It has also invested recently in Inkling, which makes interactive textbooks for the iPad. Born in India, he joined Netscape in 1994. His next company, Junglee, was bought by Amazon in 1998.
The 53-year-old was elected Stanford University trustee last December.
Wadhwani is founder, Chairman & CEO of Symphony Technology Group, a private equity firm investing in software and software services company.
An IIT Mumbai alumnus, Wadhwani moved to the US in 1969 and earned a PhD in electrical engineering at Carnegie Mellon.
The 62-year-old California resident founded software firm Aspect in 1991. "Wadhwani rode tech bubble onto The Forbes 400 in 1999 with USD 9.3 billion sale of Aspect Development software firm to i2 Technologies," Forbes said.
Silicon Valley venture capitalist Khosla co-founded Sun Microsystems in 1982. "Khosla had a very good year. SKS, the Indian microfinance lender he backed in 2006, went public in August; his personal stake was worth a recent USD 90 million," Forbes said.
He joined Sun investor John Doerr at venture firm Kleiner Perkins Caufield & Byers in 1996 and started Khosla Ventures in 2004. His firm raised USD 1.1 billion in 2009, the most by a venture firm in 3 years.
An "avid-skydiver and hand glider," the 55-year-old is an IIT alumnus and his Three Dog Vineyards, named after his dogs, produces a thousand bottles of cabernet sauvignon a year.
The list also includes the likes of industrialist Warren Buffett, Chief Executive Officer of Berkshire Hathaway Inc and Gates', Oracle's co-founder and CEO Larry Ellison, Facebook founder and CEO Mark Zuckerberg, Apple's boss Steve Jobs and media mogul Oprah Winfrey.
Forbes said the net worth of its rich list climbed 8 per cent this year to $1.4 trillion compared to $1.27 trillion last year.
Wealth rose for 217 members of the list, while 85 saw a decline. It also said that 16 new members joined the list this year, courtesy "inheritance, appreciation and big buyouts."
However, 34 people fell off the list in 2010, including Raj Rajaratnam, founder of the Galleon Group hedge fund, who is facing 185 years in prison.
The top 10 on the list gained 24.9 billion dollars.
The price of admission to the list is back up to $one billion from 2009 when $950 million was enough to make it to the top 400.